CN104870017A

CN104870017A - Pharmaceutical compositions containing a pde4 inhibitor and a pi3 delta or dual pi3 delta-gamma kinase inhibitor

Info

Publication number: CN104870017A
Application number: CN201380063816.2A
Authority: CN
Inventors: 斯瓦鲁普·库马尔·文卡塔·萨蒂亚·瓦卡拉恩卡; 斯里坎坦·维斯瓦纳德哈
Original assignee: Rhizen Pharmaceuticals SA
Current assignee: Rhizen Pharmaceuticals SA
Priority date: 2012-11-08
Filing date: 2013-11-07
Publication date: 2015-08-26
Anticipated expiration: 2033-11-07
Also published as: WO2014072937A1; US10058539B2; KR20150079745A; CA2889905A1; JP7030656B2; US20150272936A1; EP2916868A1; HK1214159A1; JP2015536973A; EA201590721A1; CN111904962A; US10413532B2; JP6434416B2; KR20210010958A; US20200237735A1; CN104870017B; US20190083475A1; US20170319558A1; US9737521B2; US11065236B2

Abstract

This present invention relates to a method of treating a autoimmune, respiratory and/or inflammatory disease or condition (e.g., psoriasis, rheumatoid arthritis, asthma, COPD). The method comprises administering a PI3K Delta inhibitor or a dual PI3K Delta- Gamma inhibitor and a PDE4 inhibitor. The present invention also relates to pharmaceutical compositions containing a PI3K Delta or dual PI3K Delta-Gamma inhibitor and a PDE4 inhibitor.

Description

Pharmaceutical composition containing PDE4 inhibitor and PI3 δ or dual PI3 δ-γ inhibitors of kinases

This application claims the rights and interests of the 4688/CHE/2012 that the Indian patent application 2762/CHE/2012 and 2012 that submits on November 8th, 2012 submits to 8, on November, it is respectively incorporated to way of reference entirety naturally hereby.

Technical field

The present invention relates to the method that one treats autoimmune, respiratory and/or inflammatory diseases or condition of illness (such as psoriasis, rheumatoid arthritis, asthma and COPD), it is by using PI3K δ inhibitor or dual PI3K δ-gamma inhibitors and PDE4 inhibitor.The invention still further relates to the pharmaceutical composition containing PI3K δ or dual PI3K δ-gamma inhibitors and PDE4 inhibitor.

Background technology

Autoimmune, respiratory and inflammatory diseases; such as rheumatoid arthritis (RA), psoriasis, systemic lupus erythematosus (SLE), chronic obstructive pulmonary disease (COPD) and asthma, be regulate and control with immune system that exception or hyperkinesia be associated chronic and frequent progressive disease.The cause of these diseases and facilitate factor still indefinite.It is characterized in that the complex cell between congenital and the multiple inflammatory cell of adaptive immune system interacts.Therefore, the heterogeneity of the D Ety of these condition of illness and complexity make to find new cell target and have more challenge, and contrast " an innocent person " onlooker because do not know in cellular infiltration, who is only main pathology and works factor.

Rheumatoid arthritis (RA) is a kind of Progressive symmetric erythrokeratodermia, general autoimmune disease, it is characterized in that multiple joints chronic inflammation, with the systemic symptoms be associated, such as tired.This inflammation causes arthralgia, stiff and swelling, thus causes function of joint to be lost because of bone and cartilage destruction, often can cause progressive disability.The probability suffering from other systemic complications of patient evolution of RA increases, described complication such as osteoporosis, anemia and other affect the disease of lung and skin.Described disease also affects average life expectancy, makes the lost of life 3 to 7 years.

Multiple treatment is had to be available for managing RA.Some solve sign and the symptom of RA, and the object of other treatment is to change the course of disease and the systemic effect affecting RA energetically, such as tired and anemia.

Current treatments comprises use:

Biological preparation: these biological preparation are genetically engineered medicines, its target specific cells surface marker or be called messenger substances in the immune system of cytokine, described cytokine is produced by cell to regulate and control other cell in inflammatory response process.The example of the specific cytokines of biological preparation targeting is tumor necrosis factor α (TNF α).

Tradition Disease Modifying Anti-Rheumatic medicine (DMARD): these medicines are nonspecific immunity compacting medicines, are intended to sign and the symptom of resisting RA, and slow down Progressive symmetric erythrokeratodermia destruction of joint.These are treated frequent combination with one another and use, or combination biological agent uses, to improve patient's response.

Glucocorticoid (corticosteroid): these are anti-inflammatory drugs relevant with hydrocortisone, the steroid that hydrocortisone being natural generation in a kind of body, working by offsetting inflammation.But the side effect of glucocorticoid, comprises hyperglycemia, osteoporosis, hypertension, body weight growth, cataract, sleeping problems, muscle loss and the susceptibility to infection, limits it and uses.

Non-steroid anti-inflammatory medicine (NSAID): the sign of these medication managements RA and symptom, such as reduces pain, swelling and inflammation, but does not change the course of disease or slow down the carrying out of destruction of joint.

Also there is the RA therapy of multiple targeting other component immune.These therapies comprise the biological treatment that targeting contributes to reducing the replaceability cytokine such as interleukin-6 (IL-6) that joint and systemic inflammation and RA are carried out.

Asthma is modal chronic disease in child, and affects millions of adults.The whole world has about 23.5 thousand ten thousand people to suffer from this disease.But the cause of asthma is not yet fully understood.

Chronic obstructive pulmonary disease (COPD) is a kind of very general condition of illness, and is the main cause of whole world M & M.Along with disease is carried out, the patient of suffering from copd may trend towards frequent aggravation, thus causes patient anxiety, health status deterioration, decline in pulmonary function and mortality rate to increase.These events that respiratory function is worsened cause health care to utilize, are in hospital and cost increase.Worse, frequent aggravation is associated with declining more fast of pulmonary function, and then shortens life expectancy.

According to the recommendation of chronic obstructive pulmonary disease whole world proposal (GOLD), a gamma therapy of COPD is long acting beta-2-agonists, long-acting muscarine antagonist and induction type corticosteroid.But these medicines reduce the symptom and aggravation that join with disease association, instead of its molecule of targeting and cell base.Therefore, the further improvement to COPD therapy is still needed.

PI-3 kinase (PI3K) belongs to a class lipid within endothelial cells kinases, and it makes 3 di of the inositol ring of lipositol (PI), thus produces lipid second message,second messenger.There are four kinds of I class PI3K isoforms---α, β, δ and γ.

IC87114 (2-((6-amino-9H-purine-9-base) methyl)-5-methyl-3-o-tolyl quinazoline-4 (3H)-one) is species specificity PI3K δ inhibitor (Sadhu, J.Immunology, 1; 170 (5): 2647-2654,2003; International publication number WO 2010/111432 and U.S. Publication No 2010/0249155 and 2010/0168139).

CAL-101 (ending for Larry this (Idelalisib)), TGR-1202, AMG-319 and INCB040093 have been reported as the inhibitor of PI3K δ and have been in positive clinical development.IPI-145 (Du Weilisi (duvelisib)) and CAL130 has been reported as the double inhibitor of PI3K δ/γ, and IPI-145 is in for the clinical research of cancer and asthma and combination methotrexate in the clinical research of RA.

Phosphodiesterase-4 (PDE4) suppresses to be a kind of approach that COPD treats.Roflumilast (Roflumilast), a kind of Novel PDE 4 inhibitors, it reduces the airways inflammation in COPD, as by expectorant neutrophil cell and eosinophil counting evaluate.But roflumilast shows dose-dependent toxicity, which has limited the use of roflumilast under higher dosage.Calverley, P, Rabe K, waits people, Roflumilast in Symptomatic ChronicObstructive Pulmonary Disease:Two Randomized Clinical Trials.TheLancet 2009; 374:685-694; And Fabbri, L, Calverley, P, Deng people, Roflumilastin Moderate to Severe Chronic Obstructive Pulmonary Disease Treatedwith Longacting Bronchodilators:Two Randomized Clinical Trials.TheLancet 2009; 374:695-703.

In addition, Celgene demonstrates positive findings in studying for two III phases of PDE-4 inhibitor A Pusite (aprelimilast) in treatment arthritic psoriasis.

Another kind of PDE4 inhibitor is the AN2728 from Anacor Pharmaceuticals, and its II phase completed for atopic dermatitis is studied.Recently, Chiesi Group announces for successfully completing to test the I phase of CHF6001, and CHF6001 is a kind of suction-type PDE4 inhibitor, and it is developed for treatment inflammatory respiratory disorder, such as chronic obstructive pulmonary disease (COPD) and asthma.

But with regard to effect and toxicity, PDE4 inhibitor has narrow treatment window.

Although current available Intervention Therapy, still need the therapeutic treatment of the improvement for autoimmune disorder such as RA and psoriasis and respiratory disorder such as asthma and COPD.

Therefore, new pharmaceutical composition that the object of this invention is to provide increased activity, that be used for the treatment of respiratory and/or inflammatory diseases and condition of illness and method.

Summary of the invention

The present invention relates to the method that one treats autoimmune, respiratory and/or inflammatory diseases or condition of illness such as rheumatoid arthritis (RA) or chronic obstructive pulmonary disease (COPD).Described method comprises the combination of using the following: (i) PI3K δ or dual PI3K δ and gamma inhibitors; And (ii) PDE4 inhibitor.Surprisingly find PI3K δ or dual PI3K δ and gamma inhibitors and PDE-4 inhibitor synergism (that is, described combination show activity be significantly greater than the activity of will expect based on PI3K δ or dual PI3K δ and gamma inhibitors and the respective separately indivedual activity of PDE-4 inhibitor).PI3K δ or dual PI3K δ and gamma inhibitors and PDE4 inhibitor can be used jointly (such as, by having both in single dosage form, or simultaneously or the independent dosage form of continuous administration).This combination is particularly useful for treatment asthma, allergic rhinitis, non-allergic rhinitis, RA, COPD and atopic dermatitis.

An embodiment is the method that one treats autoimmune, respiratory and/or inflammatory diseases or condition of illness (such as RA or COPD), and it comprises uses PI3K δ inhibitor and PDE4 inhibitor to patient in need.Preferably, the PI3K δ inhibitor of administering therapeutic effective dose and PDE4 inhibitor.

Another embodiment is the method that one treats autoimmune, respiratory and/or inflammatory diseases or condition of illness (such as RA or COPD), and it comprises uses dual PI3K δ and gamma inhibitors and PDE4 inhibitor to patient in need.Preferably, the dual PI3K δ of administering therapeutic effective dose and gamma inhibitors and PDE4 inhibitor.

Another embodiment is a kind of pharmaceutical composition, and it comprises (i) PI3K δ or dual PI3K δ and gamma inhibitors; And (ii) PDE4 inhibitor.Described pharmaceutical composition can be used for treatment autoimmune, respiratory and inflammatory diseases and condition of illness, such as treats RA and COPD.

In one embodiment, pharmaceutical composition comprises PI3K δ inhibitor and PDE4 inhibitor.In another embodiment, pharmaceutical composition comprises dual PI3K δ and gamma inhibitors and PDE4 inhibitor.In a preferred embodiment, pharmaceutical composition comprises the PI3K δ or dual PI3K δ and gamma inhibitors and PDE4 inhibitor that treat effective dose.

The activating agent of method and composition permission small amount as herein described treats autoimmune, respiratory and/or inflammatory diseases and condition of illness, and then allows cost savings, reduces side effect, and allows to continue long period section in a more effective manner and treat.

In one embodiment, PI3K δ or dual PI3K δ and gamma inhibitors are formula (I) compounds:

Or its tautomer, its N-oxide, its pharmaceutically acceptable ester, its prodrug or its pharmaceutically acceptable salt, wherein:

The R of each appearance is independently selected from hydrogen, halogen ,-OR ^a, CN, replacement or unsubstituted C _1-6alkyl, replacement or unsubstituted C _2-6thiazolinyl, replacement or unsubstituted C _2-6alkynyl, replacement or unsubstituted C _3-8cycloalkyl and replacement or unsubstituted heterocyclic group;

R ¹and R ²can be identical or different, and independently selected from hydrogen, halogen and replacement or unsubstituted C _1-6alkyl, or be all directly bonded to monatomic R ¹and R ²can engage to be formed oxo group (=O) or replace or unsubstituted, saturated or undersaturated 3-10 person's ring (comprise R ¹and R ²in conjunction with carbon atom), described ring optionally comprises and one or morely may be the same or different and be selected from O, NR ^aand the hetero atom of S;

Cy ¹for being selected from following monocyclic groups: replacement or unsubstituted cycloalkyl, replacement or unsubstituted heterocyclic group, replacement or unsubstituted aryl and replacement or unsubstituted heteroaryl;

Cy ²be selected from replacement or unsubstituted heterocyclic group, replacement or unsubstituted aryl and replacement or unsubstituted heteroaryl;

L ₁do not exist or be selected from-(CR ^ar ^b) _q-,-O-,-S (=O) _q-,-NR ^a-or-C (=Y)-;

The R of each appearance ^aand R ^bto may be the same or different and independently selected from hydrogen, halogen, hydroxyl, cyano group, replacement or unsubstituted (C _1-6) alkyl ,-NR ^cr ^d(wherein R ^cand R ^dbe hydrogen, halogen, hydroxyl, cyano group, replacement or unsubstituted (C independently _1-6) alkyl and (C _1-6) alkoxyl) and-OR ^c(wherein R ^cfor that replace or unsubstituted (C _1-6) alkyl), or work as R ^aand R ^bdirectly be bonded to the same atomic time, its can engage to be formed oxo group (=O) or formed replace or unsubstituted, saturated or undersaturated 3-10 person's ring (comprise R ^aand R ^bthe described same atom of direct combination), described ring optionally comprises and one or morely may be the same or different and be selected from O, NR ^d(wherein R ^dfor hydrogen or replacement or unsubstituted (C _1-6) alkyl) or the hetero atom of S;

Y is selected from O, S and NR ^a;

N is 1,2,3 or 4; And

Q is 0,1 or 2.

In one embodiment, formula (I) compound is formula (II) compound:

Or its tautomer, its N-oxide, its pharmaceutically acceptable ester, its prodrug or its pharmaceutically acceptable salt, wherein R, R ¹, R ², L ₁, Cy ¹and Cy ²as above for formula (I) define.

In further embodiment, formula (I) compound is selected from formula (IA-I), (IA-II), (IA-III) and (IA-IV) compound:

R, Cy ¹, R ¹, R ², n and q as defined above;

The X of each appearance is independently selected from CR ³or N; And

The R of each appearance ³independently selected from hydrogen, hydroxyl, halogen, carboxyl, cyano group, nitro, replace or unsubstituted alkyl, replace or unsubstituted alkoxyl, replace or unsubstituted thiazolinyl, replace or unsubstituted alkynyl, replace or unsubstituted aryl, replace or unsubstituted aryl alkyl, replace or unsubstituted cycloalkyl, replace or unsubstituted cycloalkyl-alkyl, replace or unsubstituted cycloalkenyl alkyl, replace or unsubstituted cycloalkenyl group, replace or unsubstituted heteroaryl, replace or unsubstituted heteroaryl alkyl, replace or unsubstituted heterocycle, the cycloheteroalkylalkyl ring replaced, replace or unsubstituted guanidine,-COOR ^x,-C (O) R ^x,-C (S) R ^x,-C (O) NR ^xr ^y,-C (O) ONR ^xr ^y,-NR ^yr ^z,-NR ^xcONR ^yr ^z,-N (R ^x) SOR ^y,-N (R ^x) SO ₂r ^y,-(=N-N (R ^x) R ^y) ,-NR ^xc (O) OR ^y,-NR ^xr ^y,-NR ^xc (O) R ^y-,-NR ^xc (S) R ^y,-NR ^xc (S) NR ^yr ^z,-SONR ^xr ^y-,-SO ₂nR ^xr ^y-,-OR ^x,-OR ^xc (O) NR ^yr ^z,-OR ^xc (O) OR ^y-,-OC (O) R ^x,-OC (O) NR ^xr ^y,-R ^xnR ^yc (O) R ^z,-R ^xoR ^y,-R ^xc (O) OR ^y,-R ^xc (O) NR ^yr ^z,-R ^xc (O) R ^x,-R ^xoC (O) R ^y,-SR ^x,-SOR ^x,-SO ₂r ^xand-ONO ₂, the R wherein in above each group ^x, R ^yand R ^zcan be hydrogen, replace or unsubstituted alkyl, replace or unsubstituted alkoxyl, replace or unsubstituted thiazolinyl, replace or unsubstituted alkynyl, replace or unsubstituted aryl, replace or unsubstituted aryl alkyl, replace or unsubstituted cycloalkyl, replace or unsubstituted cycloalkyl-alkyl, replace or unsubstituted cycloalkenyl group, replace or unsubstituted heteroaryl, replace or unsubstituted heteroaryl alkyl, replace or unsubstituted heterocycle, that replace or unsubstituted cycloheteroalkylalkyl ring or replacement or unsubstituted amino, or R ^x, R ^yand R ^zin arbitrarily both can engage to form replacement or unsubstituted saturated or undersaturated 3-10 person's ring, described ring optionally comprises and may be the same or different and be selected from O, NR ^f(wherein R ^ffor hydrogen or replacement or unsubstituted alkyl) or the hetero atom of S.

In preferred embodiments, the present invention relates to the method for the treatment of autoimmune, respiratory and/or inflammatory diseases or condition of illness, it comprises uses PI3K δ or dual, the combination of PI3K δ and gamma inhibitors and PDE4 inhibitor, wherein PI3K δ or dual PI3K δ and gamma inhibitors are the compound of formula (I), (II), (IA-I), (IA-II), (IA-III) or (IA-IV) as mentioned above.

In another preferred embodiment of the present, the present invention relates to the pharmaceutical composition comprising PI3K δ or dual PI3K δ and gamma inhibitors and PDE4 inhibitor, wherein described above, PI3K δ or dual PI3K δ and gamma inhibitors are the compound of formula (I), (II), (IA-I), (IA-II), (IA-III) or (IA-IV).

In preferred embodiments, formula (I) compound is selected from:

2-[(6-amino-9H-purine-9-base) methyl] the bromo-3-phenyl of-6--4H-benzopyran-4-one;

The bromo-2-of 6-(morpholinomethyl)-3-phenyl-4H-benzopyran-4-one;

The bromo-2-of 6-(morpholinomethyl)-3-phenyl-4H-benzopyran-4-one hydrochlorate;

2-[(6-amino-9H-purine-9-base) methyl]-3-phenyl-4H-benzopyran-4-one;

2-(morpholinomethyl)-3-phenyl-4H-benzopyran-4-one;

2-(morpholinomethyl)-3-phenyl-4H-benzopyran-4-one hydrochlorate;

2-[(1H-benzo [d] imidazoles-1-base) methyl] the bromo-3-phenyl of-6--4H-benzopyran-4-one;

The bromo-2-of 6-[(4-methyl isophthalic acid H-benzo [d] imidazoles-1-base) methyl]-3-phenyl-4H-benzopyran-4-one;

2-[(1H-benzo [d] imidazoles-1-base) methyl]-3-phenyl-4H-benzopyran-4-one;

2-[(4-methyl isophthalic acid H-benzo [d] imidazoles-1-base) methyl]-3-phenyl-4H-benzopyran-4-one;

2-[(the chloro-9H-purine of 6--9-base) methyl]-3-phenyl-4H-benzopyran-4-one;

The bromo-2-of 6-[(the chloro-9H-purine of 6--9-base) methyl]-3-phenyl-4H-benzopyran-4-one;

2-((9H-purine-6-base sulfo-) methyl)-3-phenyl-4H-benzopyran-4-one;

2-[(1H-imidazoles-1-base) methyl]-3-phenyl-4H-benzopyran-4-one;

2-[(9H-purine-6-base sulfo-) methyl] the bromo-3-phenyl of-6--4H-benzopyran-4-one;

2-((4-amino-1H-pyrazolo [3,4-d] pyrimidine-1-base) methyl) the bromo-3-phenyl of-6--4H-benzopyran-4-one;

2-[(6-amino-9H-purine-9-base) methyl] the bromo-3-of-6-(4-fluorophenyl)-4H-benzopyran-4-one;

2-[(6-amino-9H-purine-9-base) methyl]-3-(4-fluorophenyl)-4H-benzopyran-4-one;

The bromo-3-of 6-(4-fluorophenyl)-2-(morpholinomethyl)-4H-benzopyran-4-one;

The bromo-3-of 6-(4-fluorophenyl)-2-(morpholinomethyl)-4H-benzopyran-4-one hydrochlorate;

3-(4-fluorophenyl)-2-(morpholinomethyl)-4H-benzopyran-4-one;

3-(4-fluorophenyl)-2-(morpholinomethyl)-4H-benzopyran-4-one hydrochlorate;

2-[(6-amino-9H-purine-9-base) methyl] the bromo-3-o-tolyl of-6--4H-benzopyran-4-one;

7-[(the bromo-4-oxo of 6--3-phenyl-4H-.alpha.-5:6-benzopyran-2-base) methyl]-1,3-dimethyl-1H-purine-2,6 (3H, 7H)-diketone;

2-(1-(6-amino-9H-purine-9-base) ethyl) the bromo-3-phenyl of-6--4H-benzopyran-4-one;

2-(1-(9H-purine-6-base sulfo-) ethyl) the bromo-3-phenyl of-6--4H-benzopyran-4-one;

2-(1-(6-amino-9H-purine-9-base) ethyl)-3-phenyl-4H-benzopyran-4-one;

(S)-2-(1-(9H-purine-6-base is amino) ethyl) the bromo-3-phenyl of-6--4H-benzopyran-4-one;

2-((9H-purine-6-base is amino) methyl) the bromo-3-phenyl of-6--4H-benzopyran-4-one;

2-(1-(4-amino-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the bromo-3-phenyl of-6--4H-benzopyran-4-one;

2-((6-amino-9H-purine-9-base) methyl)-6-methoxyl group-3-phenyl-4H-benzopyran-4-one;

2-(1-(6-amino-9H-purine-9-base) ethyl) the bromo-3-of-6-(2-fluorophenyl)-4H-benzopyran-4-one;

2-((6-amino-9H-purine-9-base) methyl) the bromo-3-of-6-(2-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-phenyl-4H-benzopyran-4-one;

2-(1-(6-amino-9H-purine-9-base) propyl group)-3-phenyl-4H-benzopyran-4-one;

2-(1-(6-amino-9H-purine-9-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-((6-amino-9H-purine-9-base) methyl)-3-(2-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(6-amino-9H-purine-9-base) ethyl)-3-(2-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(6-amino-9H-purine-9-base) propyl group)-3-(2-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(6-amino-9H-purine-9-base) propyl group)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(6-amino-9H-purine-9-base) propyl group)-3-(4-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(6-amino-9H-purine-9-base) propyl group) the fluoro-3-phenyl of-6--4H-benzopyran-4-one;

2-(1-(6-amino-9H-purine-9-base) ethyl)-3-(4-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(6-amino-9H-purine-9-base) ethyl) the fluoro-3-phenyl of-6--4H-benzopyran-4-one;

2-(1-(4-amino-3-(3-methoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-phenyl-4H-benzopyran-4-one;

2-(1-(4-amino-3-(3-hydroxy phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-phenyl-4H-benzopyran-4-one;

2-((9H-purine-6-base is amino) methyl)-3-phenyl-4H-benzopyran-4-one;

2-(1-(6-amino-9H-purine-9-base) ethyl)-3-o-tolyl-4H-benzopyran-4-one;

2-((9H-purine-6-base is amino) methyl)-3-(2-fluorophenyl)-4H-benzopyran-4-one;

2-((9H-purine-6-base is amino) methyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

(S)-2-(1-(9H-purine-6-base is amino) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(6-amino-9H-purine-9-base) ethyl) the fluoro-3-of-6-(2-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(6-amino-9H-purine-9-base) ethyl)-3-(3,5-difluorophenyl)-4H-benzopyran-4-one;

2-(1-(6-amino-9H-purine-9-base) ethyl) the fluoro-3-of-6-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(3-methoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(3-hydroxy phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-((4-amino-3-(3-methoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) methyl)-3-phenyl-4H-benzopyran-4-one;

2-((4-amino-3-(3-hydroxy phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) methyl)-3-phenyl-4H-benzopyran-4-one;

2-((4-amino-3-(3-methoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) methyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-((4-amino-3-(3-hydroxy phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) methyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

(R)-2-(1-(9H-purine-6-base is amino) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

(S)-2-(1-(9H-purine-6-base is amino) ethyl) the fluoro-3-phenyl of-6--4H-benzopyran-4-one;

2-((4-amino-3-iodo-1H-pyrazolo [3,4-d] pyrimidine-1-base) methyl)-3-phenyl-4H-benzopyran-4-one;

2-(1-(4-amino-3-iodo-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-phenyl-4H-benzopyran-4-one;

2-(1-(4-amino-3-iodo-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-((4-amino-3-iodo-1H-pyrazolo [3,4-d] pyrimidine-1-base) methyl) the fluoro-3-phenyl of-6--4H-benzopyran-4-one;

2-(1-(4-amino-3-iodo-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-phenyl of-6--4H-benzopyran-4-one;

2-(1-(4-amino-3-iodo-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-6-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-iodo-1H-pyrazolo [3,4-d] pyrimidine-1-base) propyl group)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-((4-amino-3-(pyridin-3-yl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) methyl)-3-phenyl-4H-benzopyran-4-one;

2-((4-amino-3-(3-hydroxyl third-1-alkynyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) methyl)-3-phenyl-4H-benzopyran-4-one;

2-((4-amino-3-(1H-pyrazoles-4-base)-1H-pyrazolo [3,4-d] pyrimidine-1-base) methyl)-3-phenyl-4H-benzopyran-4-one;

2-((4-amino-3-(3-(hydroxymethyl) phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) methyl)-3-phenyl-4H-benzopyran-4-one;

2-((4-amino-3-(1H-indazole-4-base)-1H-pyrazolo [3,4-d] pyrimidine-1-base) methyl)-3-phenyl-4H-benzopyran-4-one;

2-((4-amino-3-(3-fluorophenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) methyl)-3-phenyl-4H-benzopyran-4-one;

2-((4-amino-3-(3-hydroxypropyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) methyl)-3-phenyl-4H-benzopyran-4-one;

N-(3-(4-amino-1-((4-oxo-3-phenyl-4H-.alpha.-5:6-benzopyran-2-base) methyl)-1H-pyrazolo [3,4-d] pyrimidin-3-yl) phenyl) acetamide;

2-((4-amino-3-(the fluoro-5-methoxyphenyl of 3-)-1H-pyrazolo [3,4-d] pyrimidine-1-base) methyl)-3-phenyl-4H-benzopyran-4-one;

2-((4-amino-3-(the fluoro-5-hydroxy phenyl of 3-)-1H-pyrazolo [3,4-d] pyrimidine-1-base) methyl)-3-phenyl-4H-benzopyran-4-one;

2-((4-amino-3-(the fluoro-5-methoxyphenyl of 3-)-1H-pyrazolo [3,4-d] pyrimidine-1-base) methyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-((4-amino-3-(the fluoro-5-hydroxy phenyl of 3-)-1H-pyrazolo [3,4-d] pyrimidine-1-base) methyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(1H-pyrazoles-4-base)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-phenyl-4H-benzopyran-4-one;

2-(1-(4-amino-3-(1H-indazole-6-base)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-phenyl-4H-benzopyran-4-one;

2-(1-(4-amino-3-(3-hydroxy-3-methyl fourth-1-alkynyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-phenyl-4H-benzopyran-4-one;

2-(1-(4-amino-3-(1H-pyrazoles-4-base)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

(S)-2-(1-(9H-purine-6-base is amino) ethyl)-3-phenyl-4H-benzopyran-4-one;

(S)-2-(1-(9H-purine-6-base is amino) ethyl) the fluoro-3-of-6-(3-fluorophenyl)-4H-benzopyran-4-one;

2-((4-amino-3-(1H-indazole-6-base)-1H-pyrazolo [3,4-d] pyrimidine-1-base) methyl)-3-phenyl-4H-benzopyran-4-one;

2-(1-(4-amino-3-(the fluoro-5-methoxyphenyl of 3-)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(the fluoro-5-hydroxy phenyl of 3-)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(1H-indazole-4-base)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(3,5-dimethyl-1H-pyrazoles-4-base)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(3-methyl isophthalic acid H-indazole-6-base)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(1H-indazole-6-base)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(2-(hydroxymethyl) phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(the fluoro-3-methoxyphenyl of 4-)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(the fluoro-3-hydroxy phenyl of 4-)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(3-hydroxyl third-1-alkynyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(the fluoro-4-methoxyphenyl of 3-)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(the fluoro-4-hydroxy phenyl of 3-)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(3-chloro-5-methoxyl phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(the chloro-5-hydroxy phenyl of 3-)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(3-(trifluoromethoxy) phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(4-methoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(4-hydroxy phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-((6-amino-9H-purine-9-base) methyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(the fluoro-2-methoxyphenyl of 4-)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(4-fluoro-2-hydroxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-((4-amino-3-(3-aminophenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) methyl)-3-phenyl-4H-benzopyran-4-one;

2-((4-amino-3-(3-methyl isophthalic acid H-indazole-6-base)-1H-pyrazolo [3,4-d] pyrimidine-1-base) methyl)-3-phenyl-4H-benzopyran-4-one;

2-(1-(4-amino-3-(2-aminopyrimidine-5-base)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(1H-indole-6-base)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(the chloro-3-methoxyphenyl of 4-)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(4-chloro-3-hydroxyl phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(2-chloro-5-methoxyl phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(the chloro-5-hydroxy phenyl of 2-)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(3,4-Dimethoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(3,4-dihydroxy phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-((4-amino-3-(3-methyl isophthalic acid H-indazole-6-base)-1H-pyrazolo [3,4-d] pyrimidine-1-base) methyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(1H-indole-5-base)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(3-Methyl-1H-indole-5-base)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

The methyl carbamic acid tert-butyl group-(5-(4-amino-1-(1-(3-(3-fluorophenyl)-4-oxo-4H-.alpha.-5:6-benzopyran-2-base) ethyl)-1H-pyrazolo [3,4-d] pyrimidin-3-yl) thio phenyl-2-base) ester;

2-(1-(4-amino-3-(5-(amino methyl) thio phenyl-2-base)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-((4-amino-3-(3-methyl isophthalic acid H-indazole-6-base)-1H-pyrazolo [3,4-d] pyrimidine-1-base) methyl) the fluoro-3-phenyl of-6--4H-benzopyran-4-one;

2-(1-(4-amino-3-(3-methyl isophthalic acid H-indazole-6-base)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-phenyl-4H-benzopyran-4-one;

2-(1-(4-amino-3-(3-methyl isophthalic acid H-indazole-6-base)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-phenyl of-6--4H-benzopyran-4-one;

2-(1-(4-amino-3-(3-methyl isophthalic acid H-indazole-5-base)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

N-(4-(4-amino-1-(1-(3-(3-fluorophenyl)-4-oxo-4H-.alpha.-5:6-benzopyran-2-base) ethyl)-1H-pyrazolo [3,4-d] pyrimidin-3-yl) phenyl) acetamide;

2-(1-(4-amino-3-(4-aminophenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(3-methyl isophthalic acid H-indazole-6-base)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-6-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(2,3-Dihydrobenzofuranes-5-base)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(3-ethyl-1H-indazole-6-base)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(3-Methyl-1H-indole-6-base)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(2-methoxy pyrimidine-5-base)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

4-(4-amino-1-(1-(3-(3-fluorophenyl)-4-oxo-4H-.alpha.-5:6-benzopyran-2-base) ethyl)-1H-pyrazolo [3,4-d] pyrimidin-3-yl) thiophene-2-formaldehyde;

2-(1-(4-amino-3-(5-(hydroxymethyl) thio phenyl-3-base)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(2-methyl isophthalic acid H-benzo [d] imidazoles-5-base)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(3-methyl isophthalic acid H-indazole-6-base)-1H-pyrazolo [3,4-d] pyrimidine-1-base) propyl group)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-((6-amino-9H-purine-9-base) methyl) the fluoro-3-phenyl of-6--4H-benzopyran-4-one;

2-((6-amino-9H-purine-9-base) methyl) the fluoro-3-of-6-(3-fluorophenyl)-4H-benzopyran-4-one;

2-((4-amino-3-(3-fluoro-5-methoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) methyl) the fluoro-3-of-6-(3-fluorophenyl)-4H-benzopyran-4-one;

2-((4-amino-3-(3-fluoro-5-hydroxy phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) methyl) the fluoro-3-of-6-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(3-methoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-6-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(3-hydroxy phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-6-(3-fluorophenyl)-4H-benzopyran-4-one;

2-((9H-purine-6-base is amino) methyl) the fluoro-3-of-6-(3-fluorophenyl)-4H-benzopyran-4-one;

2-((9H-purine-6-base is amino) methyl) the fluoro-3-phenyl of-6--4H-benzopyran-4-one;

(R)-2-(1-(9H-purine-6-base is amino) ethyl) the fluoro-3-of-6-(3-fluorophenyl)-4H-benzopyran-4-one;

2-((4-amino-3-(1H-pyrazoles-4-base)-1H-pyrazolo [3,4-d] pyrimidine-1-base) methyl) the fluoro-3-phenyl of-6--4H-benzopyran-4-one;

2-(1-(4-amino-3-(the fluoro-4-methoxyphenyl of 3,5-bis-)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(the fluoro-4-hydroxy phenyl of 3,5-bis-)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-((4-amino-3-(3,5-bis-fluoro-4-methoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) methyl) the fluoro-3-of-6-(3-fluorophenyl)-4H-benzopyran-4-one;

2-((4-amino-3-(3,5-bis-fluoro-4-hydroxy phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) methyl) the fluoro-3-of-6-(3-fluorophenyl)-4H-benzopyran-4-one;

(+)-2-(1-(4-amino-3-(3-methyl isophthalic acid H-indazole-6-base)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

(-)-2-(1-(4-amino-3-(3-methyl isophthalic acid H-indazole-6-base)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(3,5-Dimethoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(4-methoxyl group-3,5-3,5-dimethylphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(the fluoro-5-isopropyl phenyl of 2-)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(2,3-dihydrobenzo [b] [1,4] dioxine-6-base)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(1-benzyl-1H-pyrazoles-4-base)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(2-picoline-4-base)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(3,4-dihydro-2H-benzo [b] [1,4] benzodioxepin-7-base)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(6-morpholino pyridin-3-yl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(dibenzo [b, d] furan-4-base)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(4-(benzyloxy)-3-chlorphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(the chloro-4-isopropyl phenyl of 3-)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(3-(dimethylamino) phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(4-ethyoxyl-3-fluorophenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(4-isopropyl phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(4-(trifluoromethoxy) phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(3-(4-acetylphenyl)-4-amino-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(4-(benzyloxy) phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(4-(dimethylamino) phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(4-(methyl sulphonyl) phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(3-ethoxyl phenenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(benzo [b] thio phenyl-2-base)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(5-chlorothio benzene-2-base)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(3,5-dimethyl isoxazole-4-base)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(3-propoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(furan-2-base)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(4-ethoxyl phenenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(3-chloro-4-methoxy phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(the fluoro-4-isopropyl phenyl of 3-)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(6-fluorine pyridin-3-yl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(pyrimidine-5-base)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(3-(methoxy) phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(6-hydroxyl naphthalene-2-base)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(3-isopropyl phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(1-methyl isophthalic acid H-pyrazoles-4-base)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

The fluoro-3-of 6-(3-fluorophenyl)-2-(1-(4-Methoxyphenylamino) ethyl)-4H-benzopyran-4-one;

2-(1-(4-chloro-7H-pyrrolo-[2,3-d] pyrimidin-7-yl) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-chloro-1H-pyrazolo [3,4-b] pyridine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-chloro-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-chloro-5H-pyrrolo-[3,2-d] pyrimidine-5-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(1,3-dimethyl-1H-indazole-6-base)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(2,3-dimethyl-2H-indazole-6-base)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(6-methoxynaphthalene-2-base)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(benzo [b] thio phenyl-3-base)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(2,4-dimethoxypyridin-5-base)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(6-ethyoxyl naphthalene-2-base)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

3-(4-amino-1-(1-(3-(3-fluorophenyl)-4-oxo-4H-.alpha.-5:6-benzopyran-2-base) ethyl)-1H-pyrazolo [3,4-d] pyrimidin-3-yl)-N-cyclopropyl-phenyl Methanamide;

2-(1-(4-amino-3-(3-(morpholine-4-carbonyl) phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(3-(difluoro-methoxy) phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

5-(4-amino-1-(1-(3-(3-fluorophenyl)-4-oxo-4H-.alpha.-5:6-benzopyran-2-base) ethyl)-1H-pyrazolo [3,4-d] pyrimidin-3-yl) furan-2-formaldehyde;

(S)-2-(1-(4-amino-3-(the fluoro-4-isopropyl phenyl of 3-)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

(R)-2-(1-(4-amino-3-(the fluoro-4-isopropyl phenyl of 3-)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(3-fluoro-4-isopropyl phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-6-(3-fluorophenyl)-4H-benzopyran-4-one;

(S)-2-(1-(4-amino-3-(3-fluoro-4-isopropyl phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-6-(3-fluorophenyl)-4H-benzopyran-4-one;

(R)-2-(1-(4-amino-3-(3-fluoro-4-isopropyl phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-6-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(6-amino-9H-purine-9-base) methyl)-3-(3-fluorophenyl)-5-methoxyl group-4H-benzopyran-4-one;

2-((4-amino-3-(the fluoro-4-isopropyl phenyl of 3-)-1H-pyrazolo [3,4-d] pyrimidine-1-base) methyl)-3-(3-fluorophenyl)-5-methoxyl group-4H-benzopyran-4-one;

2-((4-amino-3-(3-fluoro-4-isopropyl phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) methyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

2-((4-amino-3-(3-fluoro-5-methoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) methyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

2-((4-amino-3-(3-fluoro-5-hydroxy phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) methyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(3-fluoro-4-isopropyl phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

(+)-2-(1-(4-amino-3-(3-fluoro-4-isopropyl phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

(-)-2-(1-(4-amino-3-(3-fluoro-4-isopropyl phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(3-methyl isophthalic acid H-indazole-6-base)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

(+)-2-(1-(4-amino-3-(3-methyl isophthalic acid H-indazole-6-base)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

(-)-2-(1-(4-amino-3-(3-methyl isophthalic acid H-indazole-6-base)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(1H-pyrazoles-4-base)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(6-amino-9H-purine-9-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(3-fluoro-4-isopropyl phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(4-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(3-fluoro-4-isopropyl phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-phenyl of-5--4H-benzopyran-4-one;

2-(1-(4-amino-3-(benzofuran-2-base)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(9H-purine-6-base is amino) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

(+)-2-(1-(4-amino-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

(-)-2-(1-(4-amino-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(4-(difluoro-methoxy)-3-fluorophenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(3-fluoro-4-isopropyl phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(1H-pyrazoles-4-base)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(the fluoro-4-of 3-(tetrahydrochysene-2H-pyrans-4-base oxygen base) phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(3-isopropyl-1H-indazole-6-base)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(the fluoro-4-of 3-(piperidin-4-yl oxygen base) phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(the fluoro-4-of 3-(2-Hydroxy-ethylamino) phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(the fluoro-4-of 3-(isopropylamino) phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(4-(dimethylamino)-3-fluorophenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(3-fluoro-4-morphlinophenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(2-methyl isophthalic acid H-benzo [d] imidazoles-5-base)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(the fluoro-4-of 3-(4-methylpiperazine-1-yl) phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-(dimethylamino)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-phenyl of-5--4H-benzopyran-4-one;

2-(1-(4-amino-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(4-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(4-(difluoro-methoxy)-3-fluorophenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(4-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(4-(difluoro-methoxy)-3-fluorophenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-phenyl of-5--4H-benzopyran-4-one;

2-(1-(4-amino-3-methyl isophthalic acid H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-ethyl-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-isopropyl-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(benzo [b] thio phenyl-2-base)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-morpholino-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(dimethylamino)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(piperidin-1-yl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(6-isopropoxypyrid-3-base)-1H-pyrazolo [3,4-d] pyrimidine-1-base ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(methyl thio)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(3-fluoro-4-isopropyl phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one 4-toluenesulfonate;

2-(1-(4-amino-3-(3-methyl isophthalic acid H-indazole-6-base)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one 4-toluenesulfonate;

2-(1-(4-amino-3-(4-(1-dibenzo-p-methyl-aza-cyclobutane-3-base oxygen base)-3-fluorophenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(the fluoro-4-of 3-(trifluoromethoxy) phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(the fluoro-4-of 3-(oxetanes-3-base oxygen base) phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(pyrrolidin-1-yl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

N-(4-(4-amino-1-(1-(the fluoro-3-of 5-(3-fluorophenyl)-4-oxo-4H-.alpha.-5:6-benzopyran-2-base) ethyl)-1H-pyrazolo [3,4-d] pyrimidin-3-yl) phenyl) isobutyramide;

2-(1-(4-amino-3-(4-isobutyl phenenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(4-isopropoxy-3-aminomethyl phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(4-(5,6-dihydro-4H-1,3-oxazine-2-base) phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

4-(4-amino-1-(1-(the fluoro-3-of 5-(3-fluorophenyl)-4-oxo-4H-.alpha.-5:6-benzopyran-2-base) ethyl)-1H-pyrazolo [3,4-d] pyrimidin-3-yl)-N-methyl benzenesulfonamide;

4-(4-amino-1-(1-(the fluoro-3-of 5-(3-fluorophenyl)-4-oxo-4H-.alpha.-5:6-benzopyran-2-base) ethyl)-1H-pyrazolo [3,4-d] pyrimidin-3-yl) the fluoro-N-isopropylbenzamide of-2-;

2-(1-(4-amino-3-(4-(5-(methylamino)-1,3,4-thiadiazoles-2-base) phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

N-(4-(4-amino-1-(1-(the fluoro-3-of 5-(3-fluorophenyl)-4-oxo-4H-.alpha.-5:6-benzopyran-2-base) ethyl)-1H-pyrazolo [3,4-d] pyrimidin-3-yl) benzyl) Methanesulfomide;

4-(4-amino-1-(1-(the fluoro-3-of 5-(3-fluorophenyl)-4-oxo-4H-.alpha.-5:6-benzopyran-2-base) ethyl)-1H-pyrazolo [3,4-d] pyrimidin-3-yl)-N-cumene sulfonamide;

4-(4-amino-1-(1-(the fluoro-3-of 5-(3-fluorophenyl)-4-oxo-4H-.alpha.-5:6-benzopyran-2-base) ethyl)-1H-pyrazolo [3,4-d] pyrimidin-3-yl)-N-cyclopropyl-phenyl sulfonamide;

2-(1-(4-amino-3-(2-isopropoxypyrimidine-5-base)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

(R)/(S)-2-(1-(4-amino-3-(3-fluoro-4-morphlinophenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

4-(4-amino-1-(1-(the fluoro-3-of 5-(3-fluorophenyl)-4-oxo-4H-.alpha.-5:6-benzopyran-2-base) ethyl)-1H-pyrazolo [3,4-d] pyrimidin-3-yl) benzsulfamide;

4-(4-amino-1-(1-(the fluoro-3-of 5-(3-fluorophenyl)-4-oxo-4H-.alpha.-5:6-benzopyran-2-base) ethyl)-1H-pyrazolo [3,4-d] pyrimidin-3-yl) thiophene-2-carboxylic acid methyl ester;

2-(1-(4-amino-3-(5-methyl thio benzene-2-base)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(1H-pyrrolo-[2,3-b] pyridine-5-base)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

4-(4-amino-1-(1-(the fluoro-3-of 5-(3-fluorophenyl)-4-oxo-4H-.alpha.-5:6-benzopyran-2-base) ethyl)-1H-pyrazolo [3,4-d] pyrimidin-3-yl)-3-fluorophenyl carbamate;

2-(1-(9H-purine-6-base is amino) propyl group) the fluoro-3-phenyl of-5--4H-benzopyran-4-one;

2-(1-(4-amino-3-(3-hydroxyl third-1-alkynyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

(S)/(R)-2-(1-(4-amino-3-(3-fluoro-4-isopropyl phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one 4-toluenesulfonate;

(+)-2-(1-(9H-purine-6-base is amino) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(4-methoxyl group-3,5-3,5-dimethylphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(4-(methoxy) phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(imidazo [1,2-a] pyridine-6-base)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

(5-(4-amino-1-(1-(the fluoro-3-of 5-(3-fluorophenyl)-4-oxo-4H-.alpha.-5:6-benzopyran-2-base) ethyl)-1H-pyrazolo [3,4-d] pyrimidin-3-yl) furan-2-base) methyl carbamic acid tert-butyl ester;

2-(1-(4-amino-3-(2,4-dimethylthiazole-5-base)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(5-(morpholinomethyl) thio phenyl-2-base)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(4-(5-amino-1,3,4-thiadiazoles-2-base) phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

(-)-2-(1-(9H-purine-6-base is amino) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(1,3-dimethyl-1H-indazole-6-base)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(2,3-dimethyl-2H-indazole-6-base)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

N-(4-(4-amino-1-(1-(the fluoro-3-of 5-(3-fluorophenyl)-4-oxo-4H-.alpha.-5:6-benzopyran-2-base) ethyl)-1H-pyrazolo [3,4-d] pyrimidin-3-yl)-2-fluorophenyl) isobutyramide;

N-(4-(4-amino-1-(1-(the fluoro-3-of 5-(3-fluorophenyl)-4-oxo-4H-.alpha.-5:6-benzopyran-2-base) ethyl)-1H-pyrazolo [3,4-d] pyrimidin-3-yl)-2-fluorophenyl) acetamide;

2-(1-(4-(dimethylamino)-3-(3-fluoro-4-isopropyl phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

The fluoro-2-of 5-(1-(3-(the fluoro-4-isopropyl phenyl of 3-)-4-(methylamino)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

The fluoro-2-of 5-(1-(3-(the fluoro-4-isopropyl phenyl of 3-)-4-morpholino-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

N-(the fluoro-4-of 2-(1-(1-(the fluoro-3-of 5-(4-fluorophenyl)-4-oxo-4H-.alpha.-5:6-benzopyran-2-base) ethyl)-4-morpholino-1H-pyrazolo [3,4-d] pyrimidin-3-yl) phenyl) isobutyramide;

N-(the fluoro-4-of 2-(1-(1-(the fluoro-3-of 5-(3-fluorophenyl)-4-oxo-4H-.alpha.-5:6-benzopyran-2-base) ethyl)-4-morpholino-1H-pyrazolo [3,4-d] pyrimidin-3-yl) phenyl) isobutyramide;

(S)/(R)-2-(1-(4-amino-3-(3-fluoro-4-isopropyl phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one sulfonate;

(S)/(R)-2-(1-(4-amino-3-(3-fluoro-4-isopropyl phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

(S)/(R)-2-(1-(4-amino-3-(3-fluoro-4-isopropyl phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one camsilate;

2-(1-(4-amino-3-(4-(difluoro-methoxy)-3-fluorophenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(1H-pyrazoles-4-base)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(3-fluoro-4-morphlinophenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-phenyl of-5--4H-benzopyran-4-one;

2-(1-(4-amino-3-(3-fluoro-4-morphlinophenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(4-fluorophenyl)-4H-benzopyran-4-one;

(S)-2-(1-(4-amino-3-(3-fluoro-4-isopropyl phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(4-fluorophenyl)-4H-benzopyran-4-one;

(R)-2-(1-(4-amino-3-(3-fluoro-4-isopropyl phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(4-fluorophenyl)-4H-benzopyran-4-one;

(S)-2-(1-(4-amino-3-(4-(difluoro-methoxy)-3-fluorophenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(4-fluorophenyl)-4H-benzopyran-4-one;

(R)-2-(1-(4-amino-3-(4-(difluoro-methoxy)-3-fluorophenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(4-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-(dimethylamino)-3-(3-fluoro-4-morphlinophenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

The fluoro-2-of 5-(1-(3-(the fluoro-4-morphlinophenyl of 3-)-4-(methylamino)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

(S)-2-(1-(4-amino-3-(3-fluoro-4-isopropyl phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-phenyl of-5--4H-benzopyran-4-one;

(R)-2-(1-(4-amino-3-(3-fluoro-4-isopropyl phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-phenyl of-5--4H-benzopyran-4-one;

(S)-2-(1-(4-amino-3-(4-(difluoro-methoxy)-3-fluorophenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-phenyl of-5--4H-benzopyran-4-one;

(R)-2-(1-(4-amino-3-(4-(difluoro-methoxy)-3-fluorophenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-phenyl of-5--4H-benzopyran-4-one;

The fluoro-2-of (+)-5-(1-(3-(the fluoro-4-isopropyl phenyl of 3-)-4-(methylamino)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

The fluoro-2-of (-)-5-(1-(3-(the fluoro-4-isopropyl phenyl of 3-)-4-(methylamino)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(6-amino-2-fluoro-9H-purine-9-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(6-amino-2-fluoro-9H-purine-9-base) ethyl) the fluoro-3-of-5-(4-fluorophenyl)-4H-benzopyran-4-one;

The fluoro-3-of 5-(4-fluorophenyl)-2-(1-(6-morpholino-9H-purine-9-base) ethyl)-4H-benzopyran-4-one;

The fluoro-3-of 5-(4-fluorophenyl)-2-(1-(6-(4-methylpiperazine-1-yl)-9H-purine-9-base) ethyl)-4H-benzopyran-4-one;

2-(1-(6-(dimethylamino)-9H-purine-9-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

The fluoro-3-of 5-(3-fluorophenyl)-2-(1-(3-(3-methyl isophthalic acid H-indazole-6-base)-4-morpholino-1H-pyrazolo [34-d] pyrimidine-1-base) ethyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(3-chloro-4-morphlinophenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

(+)-2-(1-(4-amino-3-(4-isopropoxy-3-aminomethyl phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

(-)-2-(1-(4-amino-3-(4-isopropoxy-3-aminomethyl phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

(S)/(R) the fluoro-2-of-5-(1-(3-(the fluoro-4-isopropyl phenyl of 3-)-4-morpholino-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(3-chloro-4-isopropyl phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(2-methyl benzo [d] oxazole-6-base)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

The fluoro-3-of 5-(3-fluorophenyl)-2-(1-(6-morpholino-9H-purine-9-base) ethyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(the fluoro-4-isopropyl phenyl of 3-)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-3-(3-fluorophenyl)-5-morpholino-4H-benzopyran-4-one;

2-(1-(4-amino-3-(the fluoro-4-isopropyl phenyl of 3-)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-5-morpholino-3-phenyl-4H-benzopyran-4-one;

6-(4-amino-1-(1-(the fluoro-3-of 5-(3-fluorophenyl)-4-oxo-4H-.alpha.-5:6-benzopyran-2-base) ethyl)-1H-pyrazolo [3,4-d] pyrimidin-3-yl) 1-isoindolinone;

5-(4-amino-1-(1-(the fluoro-3-of 5-(3-fluorophenyl)-4-oxo-4H-.alpha.-5:6-benzopyran-2-base) ethyl)-1H-pyrazolo [3,4-d] pyrimidin-3-yl) 1-isoindolinone;

2-(1-(3-(4-acetyl group-3-fluorophenyl)-4-amino-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

The fluoro-3-of 5-(3-fluorophenyl)-2-(1-(6-(4-methylpiperazine-1-yl)-9H-purine-9-base) ethyl)-4H-benzopyran-4-one;

(S)-2-(1-(4-amino-3-(3-chloro-4-morphlinophenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

(R)-2-(1-(4-amino-3-(3-chloro-4-morphlinophenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

N-(3-(4-amino-1-(1-(the fluoro-3-of 5-(3-fluorophenyl)-4-oxo-4H-.alpha.-5:6-benzopyran-2-base) ethyl)-1H-pyrazolo [3,4-d] pyrimidin-3-yl) phenyl) Methanesulfomide;

(S)-2-(1-(6-(dimethylamino)-9H-purine-9-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

(R)-2-(1-(6-(dimethylamino)-9H-purine-9-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(9H-purine-6-base is amino) ethyl) the fluoro-3-of-5-(2-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(4-ethyoxyl-3-(trifluoromethyl) phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

2-(1-(4-amino-3-(3-fluoro-4-isopropyl phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) propyl group) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one;

(S) the fluoro-3-of-5-(3-fluorophenyl)-2-(1-(2-methoxyl group-9H-purine-6-base is amino) ethyl)-4H-benzopyran-4-one;

(R) the fluoro-3-of-5-(3-fluorophenyl)-2-(1-(2-methoxyl group-9H-purine-6-base is amino) ethyl)-4H-benzopyran-4-one;

(S)/(R) the fluoro-2-of-5-(1-(the fluoro-9H-purine of 2--6-base is amino) ethyl)-3-(3-fluorophenyl)-4H-benzopyran-4-one;

(S)/(R)-2-(1-(4-amino-3-(the fluoro-4-isopropyl phenyl of 3-)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl)-5-methyl-3-phenyl-4H-benzopyran-4-one;

2-(1-(9H-purine-6-base is amino) ethyl) the fluoro-3-o-tolyl of-5--4H-benzopyran-4-one; And

Its pharmaceutically acceptable salt.

In another preferred embodiment of the present, formula (I) compound is be selected from following PI3K δ inhibitor:

2-((6-amino-9H-purine-9-base) methyl)-5-methyl-3-o-tolyl quinazoline-4 (3H)-one (IC87114);

(S)-2-(1-((9H-purine-6-base) amino) propyl group)-5-fluoro-3-phenylquinazoline-4 (3H)-one (CAL-101, end for Larry this);

INCB040093；

AMG 319；

(S)-2-(1-(9H-purine-6-base is amino) ethyl) the fluoro-3-of-6-(3-fluorophenyl)-4H-benzopyran-4-one (Compound C);

(S)-2-(1-(4-amino-3-(3-fluoro-4-isopropyl phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-6-(3-fluorophenyl)-4H-benzopyran-4-one; And

Its pharmaceutically acceptable salt.

In another preferred embodiment of the present, formula (I) compound is be selected from following dual PI3K δ and gamma inhibitors:

(S)-3-(1-((9H-purine-6-base) is amino) ethyl)-8-chloro-2-phenyl isoquinolin quinoline-1 (2H)-one (IPI-145);

(+)-2-(1-(4-amino-3-(3-fluoro-4-isopropyl phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one (compd A 1);

(-)-2-(1-(4-amino-3-(3-fluoro-4-isopropyl phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one; And

Its pharmaceutically acceptable salt.

In preferred embodiments, PDE-4 inhibitor is A Pusite or roflumilast.

Another embodiment is a kind of test kit being used for the treatment of autoimmune, respiratory or inflammatory diseases or condition of illness, and described test kit comprises:

(i) PI3K δ or PI3K δ and gamma inhibitors; And (ii) PDE4 inhibitor, it is in single medicine compositions or independent pharmaceutical composition;

(ii) optionally, for using the instructions of PI3K δ or PI3K δ and gamma inhibitors and PDE4 inhibitor for treating autoimmune, respiratory or inflammatory diseases or condition of illness; And

(iii) optionally, for placing the container of one or more pharmaceutical compositions described.

Accompanying drawing explanation

Fig. 1 describe compd A and itself and 30 and the combination of 300nM compd B on the impact suppressing TNF α to discharge from U-937 cell.* p < 0.01 and * * * p < 0.001 (control compounds A).

The impact that the combination that Fig. 2 describes compd A and itself and 300nM compd B discharges from THP-1 cell MMP-9.

Under Fig. 3 describes to have and do not have compd B (30nM) situation, compd A is on the impact of the expression of pAkt (S473) in the differentiation U937 cell stimulated with cigarette smoke extract (CSE).

Fig. 4 describes the combination of compd A and itself and compd B (300nM) to the impact suppressing elastoser to discharge from people's neutrophil cell.* p < 0.01 and * * * p < 0.001 (control compounds A).

The combination that Fig. 5 describes compd A and itself and compd B (300nM) CSE is induced after the impact of A549 cell proliferation.* represent that p < 0.05, * * represents p < 0.01, and * * * represents p < 0.001, with administered compound A separately compared with.

Fig. 6 describes the combination of compd A and itself and compd B (300nM) to inhibitory action apoptotic in A549 cell.

Fig. 7 A, 7B and 7C describe compd A, Compound C and the inhibitory action of its combination to neutrophil infiltrates respectively.

Fig. 8 A, 8B and 8C describe compd A, compd A 1 and the inhibitory action of its combination to neutrophil infiltrates.

Fig. 9 describe compd A, Compound C and its be combined in inhibitory action to macrophages infiltration in the cellular infiltration of the acute smoke from cigarette induction of male Balb/c mice.

Figure 10 describe compd A, compd A 1 and its be combined in inhibitory action to macrophages infiltration in the cellular infiltration of the acute smoke from cigarette induction of male Balb/c mice.

Figure 11 describe compd A, Compound C and its be combined in inhibitory action to macrophages infiltration in the cellular infiltration of the chronic smoke from cigarette induction of male Balb/c mice.

Detailed description of the invention

definition

When the scope of application is herein used for physical characteristic such as molecular weight or chemical characteristic such as chemical formula, be intended to comprise all combinations of scope and sub-portfolio and specific embodiments wherein.When mentioning quantity or numerical range, it is approximation in experimental variations (or statistical experiment error) scope that term " about " means mentioned quantity or numerical range, and therefore quantity or numerical range can such as change between 1% and 15% of institute's recited number or numerical range.Term " comprises (comprising) ", and (such as " comprising (comprise) " with relational term or " comprising (comprises) " or " having (having) " or " comprising (including) ") comprises those embodiments, such as the embodiment of any formation of " being made up of described feature " or " being substantially made up of described feature " of material, compositions, method or process etc.

Following abbreviation and term have indicated implication in the whole text: PI3-K=phosphoinositide 3-kinase; PI=phosphatidylinositols; And MeI=methyl iodide.

Unless otherwise directed, otherwise abbreviation used herein have its chemistry and biological field in conventional sense.

Term " replacement or unsubstituted ", " alkyl ", " alkoxyl ", " thiazolinyl ", " alkynyl ", " aryl ", " aryl alkyl ", " cycloalkyl ", " cycloalkyl-alkyl ", " cycloalkenyl alkyl ", " cycloalkenyl group ", " heteroaryl ", " heteroaryl alkyl ", " heterocycle " (or heterocyclic radical) and " cycloheteroalkylalkyl " as international patent application no PCT/IB2010/002804 and PCT/US2012/36594 define.On the suitable pharmaceutical of PI3K inhibitor as herein described, acceptable salt comprises those described in international patent application no PCT/IB2010/002804 and PCT/US2012/36594.

Term " effective dose " or " treatment effective dose " refer to that compound as herein described or compound combination are enough to realize the application intended, the amount including but not limited to disease treatment, as defined hereinafter.Treatment effective dose can be depending on the following and becomes: the application intended (external or body in) or the experimenter treated and disease condition, the order of severity, method of application etc. of the body weight of such as experimenter and age, disease condition, it easily can be determined by those of ordinary skill in the art.Described term is also applicable in target cell, to induce particular responses, such as reduce the dosage of platelet adhesion and/or cell migration.Concrete dosage will depend on selected specific compound, dosage regimen to be observed, whether combine other compound administration, use opportunity, be applied its tissue and carry its physical delivery system and become.

As used herein, term " treatment (treatment and treating) " refers to the approach obtaining useful or results needed, and described useful or results needed includes but not limited to treatment benefit and/or prevention benefit.Treatment benefit means to eradicate or improve the potential disease for the treatment of.In addition, by eradicating or improving one or more pathophysiological condition be associated with potential disease, realize treatment benefit, although patient may still suffer from potential disease to make to observe in patients to transfer.For prevention benefit, compositions can be administered to the patient of risky development specified disease, or be administered to the patient of one or more pathophysiological condition reporting disease, even if the diagnosis to this disease not yet may be made.

" therapeutical effect " is forgiven treatment benefit as previously discussed and/or is prevented benefit as the term is employed herein.Preventive effect comprises and postpones or eliminate a disease or condition of illness occurs, postpones or eliminates a disease or the paresthesia epilepsy of condition of illness, slows down, to stop or reverse disease or condition of illness carry out, or its any combination.

Term " experimenter " or " patient " refer to animal, such as mammal, such as people.Method as herein described can be used for human therapeutics and application for animals.In some embodiments, patient is mammal, and in some embodiments, patient behaves.For object for animals, term " experimenter " and " patient " include but not limited to farming animals, comprise cattle, sheep, pig, horse and goat; Companion animals, such as Canis familiaris L. and cat; Field animal and/or zoo animal; Laboratory animal, comprises mice, rat, rabbit, Cavia porcellus and hamster; And birds, such as chicken, turkey, duck and goose.

Term " Selective depression (selective inhibition or selectively inhibit) " as being applied to bioactivator refers to compared with the signaling activity that misses the target, and medicament interacts via direct or indirect and target and optionally reduces the ability of target signaling activity.

As used herein, term " PI3-kinase delta selective depressant " generally refers to compared with other isozyme (α, β and γ) of PI3K family, more effectively suppresses the compound of the activity of PI3-kinase delta isozyme.Such as,, the compound of at least 20 times, at least 50 times, at least 100 times lower at least 10 times about the IC50 of remaining other I type PI3-kinases (that is, α, β and γ) than inhibitor about kinase whose 50% inhibition concentration of δ I type PI3-(IC50) that PI3-kinase delta selective depressant can refer to show.

As used herein, term " dual PI3-kinase delta and gamma inhibitors " generally refers to compared with other isozyme of PI3K family, more effectively suppresses the compound of the activity of PI3-kinase delta and γ isozyme.Therefore, compare with LY294002 with the such as wortmannin (wortmannin) of the conventional PI3K inhibitor for " non-selective PI3K inhibitor ", PI3-kinase delta and γ double inhibition immunomodulator compounds larger for the selectivity of PI3-kinase delta and γ.

Such as,, the compound of at least 20 times, at least 50 times, at least 100 times lower at least 10 times about the IC50 of remaining other I type PI3-kinases (that is, α and β) than inhibitor about kinase whose 50% inhibition concentration of δ and γ I type PI3-(IC50) that dual PI3-kinase delta and gamma selective inhibitor can refer to show.

Therapeutic Method of the present invention comprises the method being used for the treatment of the condition of illness be associated with inflammatory response.The feature of " inflammatory response " is rubescent, heating, swelling and pain (i.e. inflammation) and typically relates to tissue injury or destruction.Inflammatory response is generally by tissue injury or destroys the protective response of localization of drawing, and it is used for destroying, dilute or separate the tissue of (isolation) damaging reagent and damage.Inflammatory response obviously flows into leukocyte and/or leukocyte (such as neutrophil cell) chemotaxis is associated.Inflammatory response can be biological and viral by pathogenic infection, non-infectious mode such as wound or myocardial infarction or reperfusion following stroke, produces the immunne response of exotic antigen and autoimmune disease.Available method according to the present invention and the inflammatory response of compounds for treating forgive the condition of illness associated with the reacting phase of specificity system of defense and the condition of illness associated with the reacting phase of non-specific defense system.

Therapeutic Method of the present invention comprises the method being used for the treatment of the condition of illness be associated with inflammatory cell activation." inflammatory cell activation " refers to that inflammatory cell (includes but not limited to mononuclear cell, macrophage, T lymphocyte, bone-marrow-derived lymphocyte, granulocyte (polymorphonuclear leukocyte, comprise neutrophil cell, basophil and eosinophil), mastocyte, dendritic cell, langerhans cell and endotheliocyte) in by proliferative cell response stimulation (include but not limited to cytokine, antigen or autoantibody) induction carried out, Soluble mediators (includes but not limited to cytokine, oxygen-derived free radicals, enzyme, prostaglandins or vasoactive amines) generation, or it is new or increase the cell surface expression of amboceptor (including but not limited to major histocompatibility antigen or cell adhesion molecule) of quantity.It will be understood by a person skilled in the art that activation of a kind of in these phenotypes in these cells or its combination can impel inflammatory condition initial, maintain or aggravation.

" autoimmune disease " refers to the disease of any group as used herein, and wherein tissue injury is associated with body fluid or the cell-mediated response for health self component.

" anaphylaxis " disease generally refers to that any symptom, histologic lesion or the function of organization caused by allergy loses.

" arthritis " disease generally refers to any disease being characterised in that the arthritis pathological changes being attributable to Different types of etiopathogenises.

" dermatitis " generally refers to any one that be characterised in that in one of the chafing being attributable to Different types of etiopathogenises large class dermatosis.

As used herein, term " is used " jointly, " combined administration " and its equivalent are grammatically forgiven and used two or more medicaments to animal, and medicament and/or its metabolite are present in animal body all simultaneously.Jointly use to comprise and use with independently compositions simultaneously, do not using in the same time with independently compositions or using with a kind of compositions that two kinds of medicaments all exist.

As used herein, term " pharmaceutically acceptable salt " comprises the salt derived from such as following inorganic base: Li, Na, K, Ca, Mg, Fe, Cu, Zn and Mn; The salt of such as following organic base: N, N '-diacetyl ethylenediamine, glucamine, triethylamine, choline, hydroxide, dicyclohexylamine, metformin, benzyl amine, trialkylamine and thiamine; The salt of such as following chiral base: alkyl phenyl amine, glycinol and phenylqlvcinol; The salt of such as following natural amino acid: glycine, alanine, valine, leucine, isoleucine, nor-leucine, tyrosine, cystine, cysteine, methionine, proline, hydroxyproline, histidine, ornithine, lysine, arginine and serine; The compounds of this invention and alkyl halide, alkyl sulfate such as MeI and (Me) ₂sO ₄quaternary ammonium salt; The salt of such as following alpha-non-natural amino acid: as the aminoacid of D-isomer or replacement; The salt of guanidine; And the salt of the guanidine replaced, wherein substituent group is selected from nitro, amino, alkyl, thiazolinyl, alkynyl, the ammonium salt of ammonium or replacement and aluminum salt.Time suitable, salt can comprise acid-addition salts, and it is sulfate, nitrate, phosphate, perchlorate, borate, hydrohalide, acetate, tartrate, maleate, citrate, fumarate, succinate, palmitate, mesylate, benzoate, Salicylate, benzene sulfonate, Ascorbate, glycerophosphate and ketoglutarate.

pI3K δ and dual PI3K δ and gamma inhibitors

The example that can be used for PI3-kinase delta selective depressant in compositions as herein described and method and dual PI3-kinase delta and gamma inhibitors includes but not limited to CAL-101 (end for Larry this), compound: international publication number WO 2012/151525 disclosed in IPI-145 (Du Weilisi) and the following, U.S. Patent Publication number 2011/0118257 and 2012/0289496, the international patent application no PCT/IB2010/002804 that on November 3rd, 2010 submits to, the PCT/US2012/36594 that on May 4th, 2012 submits to, the U.S. Patent Application No. 13/933 that the PCT/US2013/055434 and 2013 submitted on July 2nd, 2013 submits to 2, on July, 856.The following also discloses other limiting examples: international publication number WO2001/081346, WO 2003/035075, WO 2005/113554, WO 200/113556, WO 2006/024666, WO 2008/118454, WO 2008/118455, WO2009/010530, WO 2009/064802, WO 2009/088986, WO 2009/088990, WO 2009/147189, WO 2010/005558, WO 2010/051042, WO2010/051043, WO 2010/056320, WO 2010/057048, WO 2010/092015, WO 2010/092962, WO 2010/096389, WO 2010/102958, WO2010/110685, WO 2010/110686, WO 2010/111432, WO 2010/123931, WO 2010/135014, WO 2010/136491, WO 2010/138589, WO2010/144513, WO 2010/151737, WO 2010/151740, WO 2010/151791, WO 2011/005119, WO 2011/008302, WO 2011/008487, WO2011/011550, WO 2011/012883, WO 2011/021038, WO 2011/022439, WO 2011/041399, WO 2011/041634, WO 2011/048111, WO2011/048936, WO 2011/055215, WO 2011/075268, WO 2011/075630, WO 2011/075643, WO 2011/101429, WO 2011/123751, WO2011/130342, WO 2011/156759, WO 2011/163195, WO 2012/003262, WO 2012/003264, WO 2012/003271, WO 2012/003274, WO2012/003278, WO 2012/003283, WO 2012/004299, WO 2012/007493, WO 2012/0135009, WO 2012/020762, WO 2012/021696, WO2012/032067, WO 2012/037204, WO 2012/037226, WO 2012/040634, WO 2012/044641, WO 2012/052753, WO 2012/055846, WO2012/061696, WO 2012/064973, WO 2012/068343, WO 2012/087784, WO 2012/087881, WO 2012/097000, WO 2012/107465, WO2012/116237, WO 2012/121953, WO 2012/125510, WO 2012/125629, WO 2012/126901, WO 2012/135160, WO 2012/135166, WO2012/135175, WO 2012/140419, WO 2012/146666, WO 2012/146667, WO 2012/148548, WO 2012/151525, US 2012/0220575, US2012/0238587, WO 2013/012915, WO 2013/012918, WO 2013/032591, WO 2013/033569, WO 2013/052699, WO 2013/057711, WO2013/067141, WO 2013/067306, WO 2013/071264, WO 2013/078441, WO 2013/082540, WO 2013/090725, WO 2013/116562, WO2013/132270, WO 2013/134288, WO 2013/136075 and WO2013/136076.PI3K inhibitor is described each incorporated herein by reference naturally with its these announcements prepared.

pDE-4 inhibitor

Enprofylline (enprofylline) is included but not limited to for the suitable PDE-4 inhibitor in compositions as herein described and method, theophylline, aminophylline, Oxtriphylline, A Pusite (apremilast), roflumilast, A Ruiluo (ariflo) (cilomilast (cilomilast)), appropriate Fei Site (tofimilast), pumafentrine (pumafentrine), Li meter Si Te (lirimilast), arofylline (arofylline), Ah 's azoles orchid (atizorame), Ao Ge meter Tan (oglemilastum), D-4418, Bay-198004, BY343, CP-325,366, D-4396 (Sch-351591), AWD-12-281 (GW-842470), NCS-613, CDP-840, D-4418, PD-168787, T-440, T-2585, V1 1294A, CI-1018, CDC-801, CDC-3052, D-22888, YM-58997, Z-15370, N-(the chloro-1-oxo-pyridin of 3,5-bis--4-base)-4-difluoro-methoxy-3-cyclopropyl-methoxy yl-benzamide, (-)-p-[(4aR*, 10bS*)-9-ethyoxyl-1,2,3,4,4a, 10b-six hydrogen-8-methoxyl group-2-methyl benzo [s] [1,6]-benzodiazine-6-base]-N, N-diisopropylbenzamide, (R)-(+)-1-(4-bromobenzyl)-4-[(3-cyclopentyloxy)-4-methoxyphenyl]-2-Pyrrolidone, 3-(cyclopentyloxy-4-methoxyphenyl)-1-(4-N '-[N-2-cyano-S-methyl-different sulfo-urea groups] benzyl)-2-Pyrrolidone, along [4-cyano group-4-(3-cyclopentyloxy-4-methoxyphenyl) cyclohexane extraction-1-carboxylic acid], 2-carbomethoxy-4-cyano group-4-(3-cyclo propyl methoxy-4-difluoro-methoxy phenyl) cyclohexane extraction-1-ketone, along [4-cyano group-4-(3-cyclo propyl methoxy-4-difluoro-methoxy phenyl) cyclohexane extraction-1-alcohol], acetic acid (R)-(+)-ethyl [4-(3-cyclopentyloxy-4-methoxyphenyl) pyrrolidine-2-subunit] ester, acetic acid (S)-(-)-ethyl [4-(3-cyclopentyloxy-4-methoxyphenyl) pyrrolidine-2-subunit] ester, 9 cyclopenta-5, 6-dihydro-7-ethyl-3-(2-thienyl)-9H-pyrazolo [3,4c]-1,2, 4-triazol [4, 3a] pyridine and 9-cyclopenta-5,6-dihydro-7-ethyl-3-(tert-butyl group)-9H-pyrazolo [3,4c]-1, 2, 4-triazol [4,3a] pyridine, it is optionally in racemic form, as enantiomer, diastereomer or as pharmaceutically acceptable salt, solvate or hydrate, or compound disclosed in WO 2012/016845 and WO2012/016889.

In preferred embodiments, PDE4 inhibitor is selected from theophylline, aminophylline, Oxtriphylline, roflumilast and A Pusite.In another preferred embodiment of the present, PDE4 inhibitor is roflumilast.In another embodiment, PDE4 inhibitor is A Pusite.

pharmaceutical composition

On the one hand, the invention provides the pharmaceutical composition comprising the following: PI3K δ or dual PI3K δ and γ double inhibitor and PDE4 inhibitor, and optionally one or more pharmaceutically acceptable carrier or excipient.

In one embodiment, pharmaceutical composition comprises the PI3K δ for the treatment of effective dose or the PDE4 inhibitor of dual PI3K δ and gamma inhibitors and treatment effective dose.In another embodiment, pharmaceutical composition comprises (i) PI3K δ of cooperative effective quantity or dual PI3K δ and gamma inhibitors; And (ii) PDE4 inhibitor.Such as, pharmaceutical composition can comprise about 0.1 μ g to about 2g, preferably about 1 μ g to about 1000mg, more preferably about 10 μ g to about 500mg, all 100 μ g according to appointment to the PI3K δ of about 100mg or dual PI3K δ and gamma inhibitors, and about 0.1 μ g is to about 500mg, preferably about 1 μ g to about 100mg, more preferably about 10 μ g to about 50mg, all 10 μ g according to appointment to the PDE4 inhibitor of about 10mg.

Pharmaceutical composition also can comprise one or more other active component, such as can be used for preventing and/or treating those of respiratory disease, such as β 2-agonist (such as albuterol (salbutamol), salmaterol (salmeterol) and Wei Lanteluo (vilanterol)); Corticosteroid (such as fluticasone propionate (fluticasone propionate) or fluticasone furoate, flunisolide (flunisolide), momestasone furoate (mometasone furoate), rofleponide (rofleponide) and ciclesonide (ciclesonide); And anticholinergic or muscarine antagonist (such as ipratropium bromide (ipratropium bromide), oxitropium bromide (oxytropiumbromide), tiotropium bromide (tiotropium bromide) and oxibutynin (oxybutynin)), and its combination.

Pharmaceutical carrier and/or excipient can be selected from diluent, filler, salt, disintegrating agent, binding agent, lubricant, fluidizer, wetting agent, controlled release matrix, coloring agent, flavoring agent, buffer agent, stabilizing agent, solubilizing agent and its combination.

Pharmaceutical composition of the present invention can separately or the activating agent combined administration other with one or more, described other activating agent such as above-described those.Pharmaceutical composition of the present invention or can be used together with one or more other activating agents in a sequential manner.When needing, pharmaceutical composition of the present invention and other active component can be used jointly, or both can sequentially use they are used as a combination.

Compound of the present invention and pharmaceutical composition are by using any approach of compound delivery to site of action, such as, but not limited to oral, intranasal, surface (such as percutaneous), in duodenum, parenteral (comprises intravenous, intra-arterial, intramuscular, Ink vessel transfusing, intraperitoneal or by injection or infusion), Intradermal, by in breast, in sheath, ophthalmic, after eyeball, in lung (such as aerosolized medicaments) or subcutaneous (comprising reservoir uses for long-term release, such as be embedded in splenic capsule, under brain or in cornea), Sublingual, per anum, per rectum, transvaginal or (be such as embedded in splenic capsule by Operation, under brain or in cornea) or by sucking.

Compositions can solid, semisolid, liquid or gaseous form be used, and can be maybe dried powder, such as lyophilized form.Pharmaceutical composition can so that the form of sending encapsulates, and described form comprises such as solid dosage forms, such as capsule, sachet, cachet, gelatin, paper, tablet, suppository, pill, tablet, lozenge and lozenge.The type of encapsulation generally will depend on required route of administration.As percutaneous preparation, also contain implanted extended release preparation.In a preferred embodiment, pharmaceutical composition is solid oral dosage form, such as tablet or capsule.

In another embodiment, pharmaceutical composition is applicable to sucking (such as passing through aerosolization).

The invention still further relates to the pharmaceutical composition according to arbitrary embodiment as herein described, it is used for the treatment of autoimmune, respiratory and/or inflammatory diseases and condition of illness.

Another embodiment of the present invention relates to a kind of method for the treatment of autoimmune, respiratory and/or inflammatory diseases and condition of illness, and it comprises the pharmaceutical composition according to arbitrary embodiment as herein described to experimenter's administering therapeutic effective dose in need.

Another embodiment of the present invention relates to the purposes of the pharmaceutical composition according to arbitrary embodiment as herein described, and it is for the manufacture of the medicament for treatment autoimmune of experimenter in need, respiratory and/or inflammatory diseases and condition of illness.

According in the pharmaceutical composition of arbitrary embodiment as herein described, PI3K δ inhibitor can be solvate, hydrate or the salt form with pharmaceutically acceptable acid or alkali.

According in the pharmaceutical composition of arbitrary embodiment as herein described, dual PI3K δ and gamma inhibitors can be solvate, hydrate or the salt form with pharmaceutically acceptable acid or alkali.

According in the pharmaceutical composition of arbitrary embodiment as herein described, PDE4 inhibitor can be solvate, hydrate or the salt form with pharmaceutically acceptable acid or alkali.

In one embodiment, the present invention relates to a kind of pharmaceutical composition according to arbitrary embodiment as herein described, wherein PDE4 inhibitor is roflumilast.

Another particular of the present invention relates to the pharmaceutical composition according to arbitrary embodiment as herein described, and wherein PDE4 inhibitor is A Pusite.

Another particular of the present invention relates to the pharmaceutical composition according to arbitrary embodiment as herein described, and wherein PDE4 inhibitor is theophylline.

therapeutic Method

Another embodiment of the present invention is a kind of method for the treatment of following disease: the disease (such as autoimmune disease) relevant with immune system, the disease relating to inflammation or disease (such as asthma, chronic obstructive pulmonary disease, rheumatoid arthritis, inflammatory bowel, glomerulonephritis, neuroinflammatory disorder, multiple sclerosis, uveitis and disorder of immune system), cancer or other proliferative disease, hepatic disease or disease or kidney disease or disease.Described method comprises the PI3K δ or dual PI3K δ and γ double inhibitor and PDE4 inhibitor that use effective dose.

PI3K δ or dual PI3K δ and γ double inhibitor and PDE4 inhibitor (with optionally other active component) can be incorporated in single medicine compositions and to use, or can use by independent pharmaceutical composition, these independent pharmaceutical compositions can or not used in the same time simultaneously.

The amount of often kind of compound to be administered depends on treated experimenter (such as mammal or the particularly mankind), the order of severity of disease or condition of illness, rate of application, the character of compound and the judgement of prescriber.In one embodiment, the effective dose of PI3K δ or PI3K δ and γ double inhibitor is about 0.001 to about 100mg/kg body weight/day, and such as about 1 to about 35mg/kg/ days, by single or fractionated dose.For the people of 70kg, this will be equivalent to about 0.05 to 7g/ sky, preferably about 0.05 to about 2.5g/ days.In one embodiment, the effective dose of PDE4 inhibitor is about 0.001 to about 100mg/kg body weight/day, such as about 0.1 to about 100mcg/kg/ days, by single or fractionated dose (such as about 7 to about 7000mcg/ days or about 10 to about 1000mcg/ days).The PI3K δ of effective dose or PI3K δ and γ double inhibitor and/or PDE4 inhibitor can be used by single or multiple dosage (such as twice daily or three times).

In another embodiment, the amount that PI3K δ or dual PI3K δ and gamma inhibitors and PDE4 inhibitor are about 0.01mg to about 1000mg by scope is separately used.

In another embodiment, daily about 0.05mg to the PI3K δ of about 7g or the PDE4 inhibitor of PI3K δ and γ double inhibitor and about 7 to about 7000mcg (such as about 10 to about 1000mcg).Such as, when PDE4 inhibitor is roflumilast, the daily roflumilast (preferably oral or by suck) of about 100 to about 2000mcg.In another embodiment, daily about 200,300,400,500 or the roflumilast (preferably oral or by suck) of 600mcg.

PI3K δ or PI3K δ and γ double inhibitor and PDE4 inhibitor can be oral or used by suction.In one embodiment, PI3K δ or PI3K δ and γ double inhibitor are used by suction, and PDE4 inhibitor is Orally administered.In another embodiment, PI3K δ or PI3K δ and γ double inhibitor and PDE4 inhibitor all Orally administered.In another embodiment, PI3K δ or PI3K δ and γ double inhibitor Orally administered, and PDE4 inhibitor is used by suction.In another embodiment, PI3K δ or PI3K δ and γ double inhibitor and PDE4 inhibitor are all used by suction.

In the other embodiment of either method as herein described, PI3K δ or dual PI3K δ and gamma inhibitors and PDE4 inhibitor are oral or are used by suction.Such as, PI3K δ or dual PI3K δ and gamma inhibitors can and PDE4 inhibitor is Orally administered.PI3K δ or dual PI3K δ and gamma inhibitors can and PDE4 inhibitor uses by sucking.One in PI3K δ or dual PI3K δ and gamma inhibitors is Orally administered, and PDE4 inhibitor is used by suction, or the one in PI3K δ or dual PI3K δ and gamma inhibitors is used by suction, and PDE4 inhibitor is Orally administered.

In the other embodiment of either method as herein described, PI3K δ or dual PI3K δ and gamma inhibitors and PDE4 inhibitor are used with the ratio of about 1: 100 to about 100: 1 by weight.

In one embodiment, the PI3K δ for the treatment of effective dose or dual PI3K δ and gamma inhibitors are twice daily within every three weeks, once using, and the PDE4 inhibitor for the treatment of effective dose is twice daily within every three weeks, once using.

The example carrying out the immune disorders for the treatment of by compound of the present invention includes but not limited to psoriasis, rheumatoid arthritis, vasculitis, inflammatory bowel, dermatitis, osteoarthritis, asthma, inflammatory myopathy, allergic rhinitis, vaginitis, interstitial cystitis, scleroderma, osteoporosis, eczema, allograft or xenotransplantation (organ, bone marrow, stem cell and other biological cells and tissues) transplant rejection, graft is to versus-host disease, lupus erythematosus, inflammatory diseases, type i diabetes, pulmonary fibrosis, dermatomyositis, house Glenn syndrome (Sjogren ' ssyndrome), thyroiditis (such as Hashimoto disease (Hashimoto ' s) and autoimmune thyroiditis), myasthenia gravis, autoimmune hemolytic anemia, multiple sclerosis, cystic fibrosis, chronic recurrent hepatitis, primary biliary cirrhosis, anaphylaxis conjunctivitis and atopic dermatitis.

Another embodiment of the present invention relates to a kind for the treatment of and is selected from following disease or the method for disease: respiratory disease and condition of illness, such as air flue and pulmonary disease, its with increase or the mucus that changes produce, and/or the inflammatory of air flue and/or occlusive disease, such as acute bronchitis, chronic bronchitis, chronic obstructive bronchitis (COPD), cough, emphysema, anaphylaxis or non-allergic rhinitis or sinusitis, chronic sinusitis or rhinitis, nasal polyp, chronic sinusitis, acute sinusitis, asthma, allergic bronchitis, alveolitis, peasant is sick, high response air flue, the bronchitis caused by such as antibacterial or virus or anthelmintic or fungus or protozoacide or other pathogenic infection or pneumonia, infantile asthma, bronchiectasis, pulmonary fibrosis, adult respiratory distress syndrome, bronchus and pulmonary edema, the bronchitis caused by Different Origin or pneumonia or interstitial pneumonia, described origin is such as breathed, toxicity on inhalation gas, steam, by heart failure, X-ray, radiation, the bronchitis that chemotherapy causes or pneumonia or interstitial pneumonia, the bronchitis be associated with collagenosis or pneumonia or interstitial pneumonia, described collagenosis such as lupus erythematosus, systemic scleroderma, non-pulmonary fibrosis, idiopathic pulmonary fibrosis (IPF), the interstitial lung disease of Different Origin or interstitial pneumonia, comprise asbestosis, silicosis, sarcoidosis, granulomatosis, cystic fibrosis or mucoviscidosis, or a-1 antitrypsin deficiency, or be selected from inflammatory diseases and condition of illness, the such as inflammatory diseases of gastro-intestinal tract of various origin, such as inflammatory pseudopolyp, Chron (Crohn ' s) disease, ulcerative colitis, arthritic diseases, such as rheumatoid arthritis, or mouth and nose pharynx, skin or eyes allergic inflammatory diseases, such as atopic dermatitis, seasonality and perennially chronic urticaria, the urticaria failing to understand cause and anaphylaxis conjunctivitis, and be particularly selected from asthma, anaphylaxis and non-allergic rhinitis, COPD and atopic dermatitis, it comprises the pharmaceutical composition according to arbitrary embodiment as herein described to patient therapeuticallv's effective dose in need.

Another embodiment of the present invention relates to the purposes of the pharmaceutical composition according to arbitrary embodiment as herein described, it is for the manufacture of for treatment respiratory and/or the medicament of inflammatory diseases and condition of illness, and wherein respiratory and/or inflammatory diseases or condition of illness are selected from asthma, anaphylaxis and non-allergic rhinitis, COPD and atopic dermatitis specifically.

Another embodiment of the present invention relates to the pharmaceutical composition according to arbitrary embodiment as herein described, it is used for the treatment of the medicament of respiratory and inflammatory diseases and condition of illness, and wherein respiratory and inflammatory diseases or condition of illness are selected from asthma, anaphylaxis and non-allergic rhinitis, COPD and atopic dermatitis specifically.

To further illustrate the present invention by following non-limiting example now.

Embodiment

As will be illustrated in the example below, compd A is roflumilast; Compd B is IC87114; Compound C is the embodiment 74 of international publication number WO 11/055215 (PCT/2010/002804), and compd A 1 is the embodiment 7 of international publication number WO 2012/151525 (PCT/US2012/036594).Representative embodiment uses roflumilast and A Pusite as PDE-4 inhibitor.

embodiment 1

The combination research of PI3K δ inhibitor and PDE-4 inhibitor

In these researchs, compd A is used as PDE4 inhibitor, and compd B and C are used as PI3K δ inhibitor.

The assessment of TNF α: U937 cell is seeded in 96 orifice plates by 100,000 cells/well, and with the compound incubation 30 minutes of desired concn, add 1 μ g/ml LPS afterwards.24 h before harvest supernatant, and according to the scheme that kit manufacturers (eBioscience, USA) is recommended, assess TNF α concentration by ELISA.Briefly, antibody coating Nunc Maxisorp plate is applied with the 1mg/ml TNF α of 100 μ l.Supernatant is transferred in plate, and hatches 2h at 37 DEG C.Add anti-TNF alpha and detect antibody and avidin-HRP, add tmb substrate afterwards.Absorbance is measured at 450 nm on Fluostar Omega (BMG Labtech, NC, USA).

Result: result display in FIG.10 μMs of compd As (roflumilast) and 300 or the combination of 30nM compd B (IC87114) respectively TNF α is discharged effectively reduce reach 75% 58% Emax.The remarkable reduction (p < 0.01) that TNF α secretes is observed for the combination being low to moderate 300nM compd A concentration and 300nM compd B, thus instruction is under PI3K δ inhibitor exists, roflumilast controls the regulation and control of the cytokine causing COPD to aggravate.

Quantize matrix metalloproteinase (MMP)-9: the gelatinase activity being measured MMP-9 by zymography.Protein (after hatching THP-1 cell 24h with 50ng PMA) on the gel containing 0.1% gelatin (Sigma, USA) in separation of supernatant.Gel renaturation is made, containing 10mM CaCl by hatching 30min in 2.5%Triton X-100 ₂and 0.05%ZnCl ₂substrate buffer solution (50mMTris-HCl, pH 7.5) at 37 DEG C overnight incubation, and to dye with Ku Masi light blue (0.5%).Bright zone in the blue background of gel proves that gelatinase activity exists.With each gel operation molecular weight marker (Fermentas, Lithuania).ImageJ1.42 (NIH, USA) is used to calculate band strength.

Result: result display in fig. 2.Compared with blank well, observe because induction MMP-9 release increases twice.Compd A is for the IC reducing MMP-9 ₅₀for 577nM, and compd B affects for MMP-9 release nothing separately under 300nM.But compd B (300nM) effectively strengthens the effect of compd A with the existence of compd A, or even under minimum test concentrations (0.1nM).

SDS-PAGE assesses phosphoric acid-Akt: by inducing 48h, U-937 differentiate monocytes to become macrophage with 50ng/ml PMA.Make cell carry out trypsinization and by 100, the density inoculation in 000/ hole, and stimulate 2h with CSE in starvation media.RIPA buffer is used to prepare lysate, and operated by SDS-PAGE, be transferred to pvdf membrane, and with phosphoric acid-Akt (S473) antibody (Cell signaling, USA), afterwards with anti-rabbit igg (Cellsignaling, USA) detection.ImageJ 1.42 (NIH, USA) is used to calculate band strength.

Result: result display in figure 3.Compd A and compd B suppress Akt phosphorylation with dosage-dependent manner respectively, its IC ₅₀value is respectively 1934 and 780nM.But the compd B of combination 30nM, observes the IC of compd A ₅₀(1.5nM) significantly reduce (about 1290 times), thus instruction is strengthened according to MMP-9 minimizing response.

Quantize Neutrophil elastase: obtain blood from local blood bank.Dextran sedimentation method is used to obtain neutrophil cell.Cell is seeded in 96 orifice plates by 100,000 cells/well, with fMLP (1 μM) and N-succinyl group-Ala-Ala-Ala-p-Nitraniline. (1 μM) (Sigma) process, and hatches 2h.By hatching the intensity of rear digested substrate under 405nm at Fluostar Omega (BMGLabtech, NC, USA) upper measurement absorbance measurement.

Result: result display in the diagram.Compd A suppresses the elastin laminin enzyme r e lease of fMLP induction, its IC ₅₀for 10.2nM (Emax=43%), the minimizing in the independent situation of compd B can be ignored (being 10% under 300nM).The combination of compd A and 300nM compd B causes Emax (65%) to increase, simultaneously the IC of compd A ₅₀correspondingly be reduced to 4.8nM.

Cell proliferating determining: A549 cell is seeded in 96 orifice plates by 10,000 cells/well, and hatches 72h with CSE.By assessing interpolation 100 μ gMTT and the quantitative determination cell viability of the solubility first cured (in DMSO) formed hatch 4h at 37 DEG C after.Remove culture medium and by dissolution of crystals in 100 μ lDMSO.Absorbance is measured at 450 nm on Fluostar Omega (BMG Labtech, NC, USA).

Result: result display in Figure 5.The combination of compd A and 300nM compd B causes the remarkable reverse of the antiproliferative effect to CSE, its IC ₅₀for 8.7nM.Do not wish that the combination of inventor's reasoning PDE4 inhibitor and PI3K δ inhibitor is used for protecting alveolar epithelial cells, and then minimizes the carrying out of COPD by under any particular theory restraint condition.

Cell cycle analysis: by A549 cell with 100, the density of 000 cells/well is seeded in 6 orifice plates, and stimulates 72h with CSE.After hatching, cell is fixed in 70% ethanol, and until analyze at being stored in 4 DEG C.Dye according to the description of manufacturer with Guava cell cycle reagent.Guava people's cell analysis system (Millipore, USA) is used to obtain cell cycle data.

Result: result display in figure 6.Cause apoptotic dose-dependent inhibition with compd A process.Use the combination of compd A and 300nM compd B, this decline is strengthened significantly further, thus indicates the potentially useful of this combination prevention lung damage.

In a word, determine with after LPS/CSE, the inhibitory action of TNF α, pAkt and MMP-9 in differentiation U937 macrophage.Assess the neutrophil cell manifested by the adjustment of elastase activity activity functional.Also measure the apoptotic protective effect of combination (compd A and B) to the pulmonary epithelial cells that CSE induces.Digital proof compd A (PDE4 inhibitor) and compd B (PI3K δ inhibitor) be combined in nanomolar concentration under reduce TNF α, pAkt and MMP-9, and effect greatly several times more independent than arbitrary compound.Combine the suppression also enlarged markedly Neutrophil elastase, thus the evidence being combined in the treatment benefit for the treatment of in COPD of PI3K δ inhibitor and PDE4 inhibitor is provided.

In people's whole blood (HWB), LPS induces TNF α: the HWB diluting fresh collection by culture medium, and hatch 15min with the inhibitor of desired concn.Add LPS (1 μ g/ml) and then hatch 24 hours.Collect supernatant, and use eBioscience TNF α ELISA kit to assess TNF α.Although use 1000nM Compound C or 2.5nM roflumilast can not cause separately remarkable response (TNF α secretes decline < 10%), but two kinds of compounds be combined in same concentrations under cause when using TNF α to secrete decline > 30%, increase more than 3 times compared with the effect of the effect namely combined and the Compound C used separately or roflumilast, thus indicate this to be combined in synergism in treatment inflammatory disease, particularly airways disorders, psoriasis and RA and treatment conformance.

The TNF α of LPS induction in PBMC: use Histopaque separation from the PBMC (peripheral blood lymphocytes) of whole blood by density gradient, and hatch 15 minutes with the inhibitor of desired concn.Add LPS (1 μ g/ml), and then hatch 24 hours.Collect supernatant, and use eBioscience TNF α ELISA kit to assess TNF α.TNF α is caused separately to secrete decline 20% although use 1000nM Compound C, 1.25nM roflumilast being added into 1000nM Compound C causes TNF α to secrete decline 80%, namely compared with independent with Compound C effect, effect increases 4 times, thus indicates this be combined in the synergism in treatment inflammatory disease, particularly airways disorders, psoriasis and RA and treat conformance.

Lung neutrophil cell increase disease lipopolysaccharide-induced in female Wistar rats: the exaggerating of neutrophil cell is raised and may be important with the postactivated development for few inflammatory diseases in air flue and lung and the course of disease, and described disease is Severe Asthma, COPD, cystic fibrosis and adult respiratory distress syndrome such as.Neutrophil cell facilitates the mechanism of these diseases can comprise the proteolytic enzyme of such as Neutrophil elastase and the release of oxygen-derived free radicals.During release, these reagent can cause in air flue bronchoconstriction, bronchial hyperreactivity, secretion excessively, epithelial lesions and tissue remodeling.

After quanrantine, fasting animals randomization is divided into multiple groups according to its body weight.Test compounds is prepared into the suspension in vehicle, described vehicle comprises 0.5% methylcellulose, and wherein Tween 80 is used as suspending agent.By oral gavage with the volume administered compound of 10mL/kg or vehicle.With ketamine by Animal Anesthesia, and the dosage administered compound pressing 1mg/kg is after 30 minutes, and tracheal strips uses LPS solution.After LPS instils 6 hours, by animal blood-letting under anaesthesia, and conduit is inserted trachea, and by endotracheal tube by the aliquot of 5ml heparinization PBS (1 unit/ml) by lung lavage 4 times (cumulative volume 20ml).By bronchovesicular (BAL) fluid storage at 2-8 DEG C, until measure total cell and differential blood count.By BAL flow centrifugation (500 × g, 10min), and gained cell precipitation is resuspended in 0.5ml heparinized saline.Use cell counter to measure leukocytic sum in BAL fluid or blood, and be adjusted to 1 × 10 ⁶cell/ml.Manual calculations classified counting of leucocyte.Use the centrifugal 100 microlitres of cells suspensions of cytospin 3 to prepare cytospin.Use blood staining solution to be dyeed by cytospin for differentiation, and examine under a microscope microscope slide to differentiate eosinophil according to morphological characteristic.Measure the quantity of often kind of cell type in cytospin in 300 leukocytes, and be expressed as the percentage ratio of total cell.Calculate the quantity of eosinophil in BALF.

Result: result is presented in Fig. 7 A, 7B and 7C.

The effective dose of roflumilast: roflumilast proves the dose-dependent inhibition to neutrophil infiltrates under 0.3,1,3 and 10mg/kg compared with matched group.Suppress percentage ratio to be respectively-7.89%, 43.46%, 68.02% and 92.21%, and 50% suppress (ED ₅₀) dosage is 1.8mg/kg.

The effective dose of Compound C: compared with matched group, after Orally administered Compound C, observes the dose-dependent inhibition to neutrophil infiltrates under 0.1,1,3 and 10mg/kg.Suppress percentage ratio to be respectively 14.15%, 57.76%, 56.93% and 81.55%, and 50% suppress (ED ₅₀) dosage is 1mg/kg.

Each roflumilast of ED50 dosage and the combination of Compound C: compared with matched group, roflumilast or Compound C illustrate inhibitory action neutrophil infiltrates being had to 36.18% and 36.56% respectively separately under 1.8 and 1mg/kg dosage.When by time combined to roflumilast (1.8mg/kg) and Compound C (dosage is 1mg/kg), compared with control animals, 78.20% is increased to the suppression of neutrophil infiltrates.

The cellular infiltration of acute smoke from cigarette induction in male Balb/c mice: before experiment starts, make animal adapt to 7 days.Based on body weight, by animal random assortment in different groups.1st day, by oral route, test compounds or vehicle are used to mice, and after 30 minutes, the animal of using test compounds is placed in systemic exposure case.At the 1st day to the 4th day, mice is made to be exposed to the mainstream smoke of 6 medicated cigarettes.Be exposed to the smog often propping up medicated cigarette and continue 10min (medicated cigarette at first 2 minutes complete after-flame and be with animal ventilator breathe air stream afterwards), the fresh room air of following 20min exposes.After every second medicated cigarette, carry out being interrupted for other 20min minute, be exposed to fresh room air.Control animal is made to be exposed to room air chamber.From the 1st day to the 4th day, by oral route, test compounds is used to animal.At the 5th day, after last smoke from cigarette (CS) exposes 24 hours, by animal blood-letting under anaesthesia, and conduit is inserted trachea, and by endotracheal tube by the aliquot of 0.5ml heparinization PBS (1 unit/ml) by lung lavage 4 times (cumulative volume 2ml).By collected bronchovesicular (BAL) fluid storage at 2-8 DEG C, until measure total cell and differential blood count.By BAL flow centrifugation (500 × g, 10min), and gained cell precipitation is resuspended in 0.5ml heparinized saline.Use hematimeter to measure leukocytic sum in BAL fluid and blood, and be adjusted to 1 × 10 ⁶cell/ml.Manual calculations classified counting of leucocyte.Use the centrifugal 40 microlitres of cells suspensions of cytospin 3 to prepare cytospin.Use blood staining solution to be dyeed by cytospin for differentiation, and examine under a microscope, to differentiate often kind of cell according to morphological characteristic.Measure the quantity of often kind of cell type in cytospin in 300 leukocytes, and be expressed as percentage ratio, and calculate the quantity of neutrophil cell and macrophage in each BAL fluid.

Result: result display in fig .9.

The effective dose of roflumilast: roflumilast prove compared with matched group under 1,3 and 10mg/kg to the dose-dependent inhibition of macrophages infiltration.Percentage ratio is suppressed to be respectively 22.2%, 51.00% and 69.11% and 50% suppression (ED ₅₀) dosage is 3.5mg/kg.Roflumilast prove compared with matched group under 1,3 and 10mg/kg to the inhibitory action of neutrophil infiltrates.Suppress percentage ratio to be respectively 70.85%, 73.69% and 83.01%, and consider that 50% of neutrophil infiltrates suppresses dosage (ED ₅₀) for combination research.

The effective dose of Compound C: compared with matched group, after Orally administered Compound C, observes the dose-dependent inhibition to macrophage and neutrophil infiltrates under 1,3 and 10mg/kg.The suppression percentage ratio of macrophages infiltration is respectively 34.84%, 42.09% and 61.77%, and 50% suppresses (ED ₅₀) dosage is 4.4mg/kg.Compound C prove compared with matched group under 1,3 and 10mg/kg to the inhibitory action of neutrophil infiltrates.Suppress percentage ratio to be respectively 29.06%, 62.38% and 74.25%, and 50% of neutrophil infiltrates suppress (ED ₅₀) dosage is 2.1mg/kg.

Each ED ₅₀the roflumilast of dosage and the combination of Compound C: compared with matched group, roflumilast or Compound C illustrate inhibitory action macrophages infiltration being had to 26.55% and 30.01% respectively separately under 3.5 and 3mg/kg dosage.When by time combined to roflumilast (3.5mg/kg) and Compound C (dosage is 3mg/kg), compared with control animals, 99.89% is increased to the suppression of macrophages infiltration.Compared with matched group, roflumilast or Compound C illustrate inhibitory action neutrophil infiltrates being had to 44.42% and 41.47% respectively separately under 3.5 and 3mg/kg dosage.Equally, compared with matched group, the combination of roflumilast (3.5mg/kg) and Compound C (dosage is 3mg/kg) illustrates suppression neutrophil infiltrates being had to 88.34%.

The cellular infiltration of chronic smoke from cigarette induction in male Balb/c mice: before experiment starts, make animal adapt to 7 days.Based on body weight, by animal random assortment in different groups.At the 1st day to the 11st day, mice is made to be exposed to the mainstream smoke of 4 medicated cigarettes.Be exposed to the smog often propping up medicated cigarette continue 10min (often prop up medicated cigarette at first 2 minutes complete after-flame and be with animal ventilator breathe air stream afterwards), the fresh room air of following 20min exposes.After every second medicated cigarette, carry out being interrupted for other 20min minute, be exposed to fresh room air.Control animal is made to be exposed to room air chamber.At the 6th day to the 11st day, before the whole body smog of 30min exposes, use test compounds by oral route.At the 12nd day, after last smoke from cigarette (CS) exposes 24 hours, by animal blood-letting under anaesthesia, and conduit is inserted trachea, and by endotracheal tube by the aliquot of 0.5ml heparinization PBS (1 unit/ml) by lung lavage 4 times (cumulative volume 2ml).By collected bronchovesicular (BAL) fluid storage at 2-8 DEG C, until measure total cell and differential blood count.By BAL flow centrifugation (500 × g, 10min), and gained cell precipitation is resuspended in 0.5ml heparinized saline.Use hematimeter to measure leukocytic sum in BAL fluid and blood, and be adjusted to 1 × 10 ⁶cell/ml.Manual calculations classified counting of leucocyte.Use the centrifugal 40 microlitres of cells suspensions of cytospin 3 to prepare cytospin.Use blood staining solution to be dyeed by cytospin for differentiation, and examine under a microscope, to differentiate often kind of cell according to morphological characteristic.Measure the quantity of often kind of cell type in cytospin in 300 leukocytes, and be expressed as percentage ratio, and calculate the quantity of neutrophil cell and macrophage in each BAL fluid.

Result: result display in fig. 11.

Each ED ₅₀the roflumilast of dosage and the combination of Compound C: compared with matched group, roflumilast or Compound C illustrate separately inhibitory action macrophages infiltration being had to 6.30% and-13.30% respectively under 1 and 1mg/kg dosage.When by time combined to roflumilast (1mg/kg) and Compound C (dosage is 1mg/kg), compared with control animals, 122.24% is increased to the suppression of macrophages infiltration.Compared with matched group, roflumilast or Compound C illustrate separately inhibitory action neutrophil infiltrates being had to 34.60% and 4.08% respectively under 1mg/kg and 1mg/kg dosage.Equally, compared with matched group, the combination of roflumilast (1mg/kg) and Compound C (dosage is 1mg/kg) illustrates suppression neutrophil infiltrates being had to 77.78%.

embodiment 2

The combination research of dual PI3K δ and gamma inhibitors and PDE-4 inhibitor

Compd A 1 is used to carry out this research as dual PI3K δ and gamma inhibitors.Compd A 1 shows for PI3K δ and PI3K γ enzyme, IC ₅₀value≤40nM.

In MH-S (mouse alveolar macrophages), TNF α: MH-S of LPS induction represents the mouse alveolar macrophages system of a large amount of TNF α of secretion after LPS induction.Cell is inoculated by 150,000 cells/well.To add before roflumilast 15 minutes, add 10nM compd A 1 (ultimate density).Add LPS (1 μ g/ml), and then hatch 4 hours.After 20 hours, collect supernatant, and use ELISA kit to assess TNF α.Compd A 1 suppresses TNF α with dosage-dependent manner, the important cytokine that namely COPD is involved in carrying out.Add 30nM compd A 1 to PDE4 inhibitor roflumilast and cause the IC independent with roflumilast ₅₀compare, IC ₅₀reduce hundreds of times, thus indicate this be combined in the synergism in airways disorders and treat conformance.Equally, although use 15nM compd A 1 or 130nM A Pusite not cause separately TNF α to secrete significantly to decline (about 10%), but two kinds of compound combinations under same concentrations cause decline more than 35%, this is therefore provided to be combined in synergism in the fresh and RA for the treatment of inflammatory disease, particularly airways disorders, Corii Bovis seu Bubali and treatment conformance.

In THP-1 (person monocytic cell), TNF α: THP-1 of LPS induction represents to have high endogenous pAKT level and the monocytic series secreting a large amount of TNF α after LPS induction.Cell is inoculated by 150,000 cells/well.To add before roflumilast 15 minutes, add 10nM compd A 1 (ultimate density).Add LPS (1 μ g/ml), and then hatch 4 hours.After 20 hours, collect supernatant, and use ELISA kit to assess TNF α.Compd A 1 suppresses TNF α with dosage-dependent manner, the important cytokine that namely COPD is involved in carrying out.Add 100nM compd A 1 to PDE4 inhibitor roflumilast and cause the IC independent with roflumilast ₅₀compare, IC ₅₀reduce hundreds of times, thus indicate this to be combined in the synergism in treatment airways disorders and therefore indicate its treatment conformance.

In people's whole blood (HWB), Con A+PMA induces IFN γ: the HWB diluting fresh collection by culture medium, and hatch 15 minutes with the inhibitor of desired concn.By adding concanavalin A (25 μ g/ml)+acetic acid Buddhist ripple Fructus Amomi Rotundus ester (50ng/ml) inducing cytokine release.After 20 hours, collect supernatant, and use ELISA kit to assess IFN γ.Although use 10nM compd A 1 separately not affect IFN γ and secrete, the combination of using 10nM compd A 1 and roflumilast causes the IC independent with roflumilast ₅₀compare, IC ₅₀decline 3 times, thus indicate this to be combined in the treatment conformance for the treatment of in airways disorders, psoriasis and RA.

In people's whole blood (HWB), LPS induces TNF α: the HWB diluting fresh collection by culture medium, and hatch 15 minutes with the inhibitor of desired concn.Add LPS (1 μ g/ml), and then hatch 24 hours.Collect supernatant, and use eBioscience TNF α ELISA kit to assess TNF α.Although use 100nM compd A 1 separately not affect TNF α and secrete, the combination of using 100nM compd A 1 and roflumilast causes the IC independent with roflumilast ₅₀compare, IC ₅₀reduce by more than 10 times, thus indicate this be combined in the synergism in treatment inflammatory disease, particularly airways disorders, psoriasis and RA and treat conformance.

The IFN γ that in PBMC, Con A+PMA induces: use Histopaque separation from the PBMC of whole blood by density gradient, and hatch 15 minutes with the inhibitor of desired concn.By adding concanavalin A (25 μ g/ml)+acetic acid Buddhist ripple Fructus Amomi Rotundus ester (50ng/ml) inducing cytokine release.After 20 hours, collect supernatant, and use ELISA kit to assess IFN γ.Although use 10nM compd A 1 separately not affect IFN γ and secrete, the combination adding 10nM compd A 1 and roflumilast causes the IC independent with roflumilast ₅₀compare, IC ₅₀reduce by 1.5 times, thus indicate this be combined in the synergism in treatment inflammatory disease, particularly airways disorders, psoriasis and RA and treat conformance.

The TNF α of LPS induction in PBMC: use Histopaque separation from the PBMC of whole blood by density gradient, and hatch 15 minutes with the inhibitor of desired concn.Add LPS (1 μ g/ml), and then hatch 24 hours.Collect supernatant, and use eBioscienceTNF α ELISA kit to assess TNF α.Cause separately TNF α to secrete decline 25% although use 10nM compd A 1, add 10nM compd A 1 and cause the IC independent with roflumilast with the combination of roflumilast ₅₀compare, IC ₅₀reduce by 3 times, thus indicate this to be combined in treatment conformance in treatment inflammatory disease, particularly airways disorders, psoriasis and RA.

Lung neutrophil cell increase disease lipopolysaccharide-induced in female Wistar rats: the program of lung neutrophil cell increase disease for LPS induction in female Wistar rats of carrying out as described in example 1 above with compd A 1 and roflumilast.

Result: result is presented in Fig. 8 A, 8B and 8C.

The effective dose of compd A 1: after Orally administered compd A 1, observes the dose-dependent inhibition to neutrophil infiltrates compared with matched group under 0.1,1 and 10mg/kg.Suppress percentage ratio to be respectively 17.83%, 51.76% and 70.21%, and 50% suppress (ED ₅₀) dosage is 1.3mg/kg.

Each ED ₅₀the roflumilast of dosage and the combination of compd A 1: compared with matched group, roflumilast or compd A 1 illustrate inhibitory action neutrophil infiltrates being had to 36.18% and 43.02% respectively separately under 1.8 and 1.3mg/kg dosage.When by time combined to roflumilast (1.8mg/kg) and compd A 1 (dosage is 1.3mg/kg), compared with control animals, 79.20% is increased to the suppression of neutrophil infiltrates.

The cellular infiltration of acute smoke from cigarette induction in male Balb/c mice: the program of carrying out the cellular infiltration for smoke from cigarette induction acute in male Balb/c mice as described in example 1 above with compd A 1 and roflumilast.

Result: result display in Fig. 10.

The effective dose of roflumilast: roflumilast prove compared with matched group under 1,3 and 10mg/kg to the dose-dependent inhibition of macrophages infiltration.Percentage ratio is suppressed to be respectively 22.2%, 51.00% and 69.11% and 50% suppression (ED ₅₀) dosage is 3.5mg/kg.Roflumilast prove compared with matched group under 1,3 and 10mg/kg to the inhibitory action of neutrophil infiltrates.Suppress percentage ratio to be respectively 70.85%, 73.69% and 83.01%, and consider that 50% of macrophages infiltration suppresses dosage (ED ₅₀) for combination research.

The effective dose of compd A 1: after Orally administered compd A 1, observes the dose-dependent inhibition to macrophage and neutrophil infiltrates compared with matched group under 0.1,0.3 and 1mg/kg.The suppression percentage ratio of macrophages infiltration is respectively 20.60%, 75.19% and 93.11%, and 50% suppresses (ED ₅₀) dosage is 0.20mg/kg.Compd A 1 to prove compared with matched group under 0.1,0.3 and 1mg/kg the inhibitory action of neutrophil infiltrates.Suppress percentage ratio to be respectively 14.76%, 51.31% and 112.83%, and 50% of neutrophil infiltrates suppress dosage (ED ₅₀) be 0.26mg/kg.

Each ED ₅₀the roflumilast of dosage and the combination of compd A 1: compared with matched group, roflumilast or compd A 1 illustrate inhibitory action macrophages infiltration being had to 26.55% and 31.33% respectively separately under 3.5 and 0.3mg/kg dosage.When by time combined to roflumilast (3.5mg/kg) and compd A 1 (dosage is 0.25mg/kg), compared with control animals, 65.52% is increased to the suppression of macrophages infiltration.Compared with matched group, roflumilast or compd A 1 illustrate inhibitory action neutrophil infiltrates being had to 44.42% and 36.69% respectively separately under 3.5 and 0.25mg/kg dosage.Equally, compared with matched group, the combination of roflumilast (3.5mg/kg) and compd A 1 (dosage is 0.25mg/kg) illustrates suppression neutrophil infiltrates being had to 79.14%.

Although describe invention herein with reference to particular, should understand these embodiments is only that principle of the present invention and application are described.Therefore, should understanding when not deviating from the spirit and scope of the present invention as previously discussed, multiple amendment can be carried out to illustrative embodiment, and other arrangement can be designed.Claim of enclosing is intended to limit scope of the present invention, and and then the method and structure that will contain in these claim and its equivalency range.

All publications cited in the application, patent and patent application are incorporated herein by reference, its degree just as each independent publication, patent or patent application by specific and indicate individually incorporated herein by reference.

Claims

1. treat a method for autoimmune, respiratory and/or inflammatory diseases or condition of illness, described method comprises to (i) PI3K δ inhibitor of experimenter's administering therapeutic effective dose in need or dual PI3K δ and gamma inhibitors; And (ii) PDE4 inhibitor.

2. method according to claim 1, it comprises uses PI3K δ inhibitor.

3. method according to claim 1, it comprises uses dual PI3K δ and gamma inhibitors.

4. according to the method in any one of claims 1 to 3, wherein said PI3K δ or dual PI3K δ and gamma inhibitors are formula (I) compound:

R ¹and R ²to may be the same or different and independently selected from hydrogen, halogen and replacement or unsubstituted C _1-6alkyl, or be all directly bonded to monatomic R ¹and R ²can engage to be formed oxo group (=O) or replace or unsubstituted, saturated or undersaturated 3-10 person's ring (comprise R ¹and R ²in conjunction with carbon atom), described ring optionally comprises and one or morely may be the same or different and be selected from O, NR ^aand the hetero atom of S;

Y is selected from O, S and NR ^a;

N is 1,2,3 or 4; And

Q is 0,1 or 2.

5. method according to claim 4, wherein said PI3K δ or dual PI3K δ and gamma inhibitors are formula (II) compound:

Or its tautomer, its N-oxide, its pharmaceutically acceptable ester, its prodrug or its pharmaceutically acceptable salt, wherein R, R ¹, R ², L ₁, Cy ¹and Cy ²as claim 4 define.

6. method according to claim 4, wherein said PI3K δ or dual PI3K δ and gamma inhibitors are formula (IA-I), (IA-II), (IA-III) or (IA-IV) compound:

The X of each appearance is independently selected from CR ³or N; And

7. the method according to any one of claim 1,2 and 4 to 6, wherein said PI3K δ inhibitor is selected from the group of the following composition:

(S)-2-(1-((9H-purine-6-base) is amino) propyl group)-5-fluoro-3-phenylquinazoline-4 (3H)-one (CAL-101);

(S)-2-(1-(4-amino-3-(3-fluoro-4-isopropyl phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-6-(3-fluorophenyl)-4H-benzopyran-4-one,

And its pharmaceutically acceptable salt.

8. the method according to any one of claim 1 and 3 to 6, wherein said dual PI3K δ and gamma inhibitors are selected from the group of the following composition:

(-)-2-(1-(4-amino-3-(3-fluoro-4-isopropyl phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) ethyl) the fluoro-3-of-5-(3-fluorophenyl)-4H-benzopyran-4-one,

And its pharmaceutically acceptable salt.

9. method according to any one of claim 1 to 8, wherein said PDE-4 inhibitor is selected from the group of the following composition: enprofylline, theophylline, aminophylline, Oxtriphylline, A Pusite, roflumilast, cilomilast, appropriate Fei Site, pumafentrine, Li meter Si Te, arofylline, Ah 's azoles is blue, Ao Ge meter Tan, D-4418, Bay-198004, BY343, CP-325,366, D-4396 (Sch-351591), AWD-12-281 (GW-842470), NCS-613, CDP-840, D-4418, PD-168787, T-440, T-2585, V 1 1294A, CI-1018, CDC-801, CDC-3052, D-22888, YM-58997, Z-15370, N-(the chloro-1-oxo-pyridin of 3,5-bis--4-base)-4-difluoro-methoxy-3-cyclopropyl-methoxy yl-benzamide, (-)-p-[(4aR*, 10bS*)-9-ethyoxyl-1,2,3,4,4a, 10b-six hydrogen-8-methoxyl group-2-methyl benzo [s] [1,6]-benzodiazine-6-base]-N, N-diisopropylbenzamide, (R)-(+)-1-(4-bromobenzyl)-4-[(3-cyclopentyloxy)-4-methoxyphenyl]-2-Pyrrolidone, 3-(cyclopentyloxy-4-methoxyphenyl)-1-(4-N '-[N-2-cyano-S-methyl-different sulfo-urea groups] benzyl)-2-Pyrrolidone, along [4-cyano group-4-(3-cyclopentyloxy-4-methoxyphenyl) cyclohexane extraction-1-carboxylic acid], 2-carbomethoxy-4-cyano group-4-(3-cyclo propyl methoxy-4-difluoro-methoxy phenyl) cyclohexane extraction-1-ketone, along [4-cyano group-4-(3-cyclo propyl methoxy-4-difluoro-methoxy phenyl) cyclohexane extraction-1-alcohol], acetic acid (R)-(+)-ethyl [4-(3-cyclopentyloxy-4-methoxyphenyl) pyrrolidine-2-subunit] ester, acetic acid (S)-(-)-ethyl [4-(3-cyclopentyloxy-4-methoxyphenyl) pyrrolidine-2-subunit] ester, 9 cyclopenta-5,6-dihydro-7-ethyl-3-(2-thienyl)-9H-pyrazolo [3,4c]-1,2,4-triazol [4,3a] pyridine and 9-cyclopenta-5,6-dihydro-7-ethyl-3-(tert-butyl group)-9H-pyrazolo [3,4c]-1,2,4-triazol [4,3a] pyridine, and its pharmaceutically acceptable salt.

10. method according to any one of claim 1 to 9, wherein said PDE-4 inhibitor is selected from the group of the following composition: theophylline, aminophylline, Oxtriphylline, roflumilast, A Pusite and its pharmaceutically acceptable salt.

11. methods according to any one of claim 1 to 10, wherein use the described PI3K δ of described treatment effective dose or the PDE4 inhibitor of dual PI3K δ and gamma inhibitors and described treatment effective dose as combination preparation simultaneously.

12. methods according to any one of claim 1 to 11, wherein order uses the PI3K δ of described treatment effective dose or the PDE4 inhibitor of dual PI3K δ and gamma inhibitors and described treatment effective dose.

13. methods according to claim 12, wherein used the PDE-4 inhibitor of described treatment effective dose before the PI3K δ of described treatment effective dose or dual PI3K δ and gamma inhibitors.

14. methods according to any one of claim 1 to 13, the PI3K δ of wherein said treatment effective dose or dual PI3K δ and gamma inhibitors are twice daily within every three weeks, once using, and the described PDE4 inhibitor of described treatment effective dose is twice daily within every three weeks, once using.

15. methods according to any one of claim 1 to 14, wherein said autoimmune, respiratory and/or inflammatory diseases or condition of illness are selected from the group of the following composition: asthma, chronic obstructive pulmonary disease, rheumatoid arthritis, inflammatory bowel, glomerulonephritis, neuroinflammatory disorder, multiple sclerosis, uveitis, psoriasis, arthritis, vasculitis, dermatitis, osteoarthritis, inflammatory myopathy, allergic rhinitis, vaginitis, interstitial cystitis, scleroderma, osteoporosis, eczema, allograft or xenotransplantation (organ, bone marrow, stem cell and other biological cells and tissues) transplant rejection, graft is to versus-host disease, lupus erythematosus, inflammatory diseases, type i diabetes, pulmonary fibrosis, dermatomyositis, house Glenn syndrome, thyroiditis, myasthenia gravis, autoimmune hemolytic anemia, cystic fibrosis, chronic recurrent hepatitis, primary biliary cirrhosis, anaphylaxis conjunctivitis, atopic dermatitis and its combination.

16. methods according to any one of claim 1 to 14, wherein said autoimmune, respiratory and/or inflammatory diseases or condition of illness are selected from the group of the following composition: asthma, allergic rhinitis, non-allergic rhinitis, rheumatoid arthritis, chronic obstructive pulmonary disease and atopic dermatitis.

17. methods according to any one of claim 1 to 16, the amount that wherein said PI3K δ or dual PI3K δ and gamma inhibitors and described PDE4 inhibitor are about 0.01mg to about 1000mg by scope is separately used.

18. methods according to any one of claim 1 to 17, wherein said PI3K δ or dual PI3K δ and gamma inhibitors and described PDE4 inhibitor are used with the ratio of about 1: 100 to about 100: 1 by weight.

19. 1 kinds of pharmaceutical compositions, it comprises (i) PI3K δ or dual PI3K δ and gamma inhibitors or its pharmaceutically acceptable salt; (ii) PDE4 inhibitor; And (iii) optionally pharmaceutically acceptable carrier, fluidizer, diluent or excipient.

20. pharmaceutical compositions according to claim 19, wherein said PI3K δ or dual PI3K δ and gamma inhibitors are formula (I) compound:

Y is selected from O, S and NR ^a;

N is 1,2,3 or 4; And

Q is 0,1 or 2.

21. pharmaceutical compositions according to claim 20, wherein said formula (I) compound is formula (II) compound:

Or its tautomer, its N-oxide, its pharmaceutically acceptable ester, its prodrug or its pharmaceutically acceptable salt, wherein R, R ¹, R ², L ₁, Cy ¹and Cy ²as claim 20 define.

22. pharmaceutical compositions according to claim 20, wherein said formula (I) compound is formula (IA-I), (IA-II), (IA-III) or (IA-IV) compound:

The X of each appearance is independently selected from CR ³or N; And

The R of each appearance ³independently selected from hydrogen, hydroxyl, halogen, carboxyl, cyano group, nitro, replace or unsubstituted alkyl, replace or unsubstituted alkoxyl, replace or unsubstituted thiazolinyl, replace or unsubstituted alkynyl, replace or unsubstituted aryl, replace or unsubstituted aryl alkyl, replace or unsubstituted cycloalkyl, replace or unsubstituted cycloalkyl-alkyl, replace or unsubstituted cycloalkenyl alkyl, replace or unsubstituted cycloalkenyl group, replace or unsubstituted heteroaryl, replace or unsubstituted heteroaryl alkyl, replace or unsubstituted heterocycle, the cycloheteroalkylalkyl ring replaced, replace or unsubstituted guanidine,-COOR ^x,-C (O) R ^x,-C (S) R ^x,-C (O) NR ^xr ^y,-C (O) ONR ^xr ^y,-NR ^yr ^z,-NR ^xcONR ^yr ^z,-N (R ^x) SOR ^y,-N (R ^x) SO ₂r ^y,-(=N-N (R ^x) R ^y) ,-NR ^xc (O) OR ^y,-NR ^xr ^y,-NR ^xc (O) R ^y-,-NR ^xc (S) R ^y,-NR ^xc (S) NR ^yr ^z,-SONR ^xr ^y-,-SO ₂nR ^xr ^y-,-OR ^x,-OR ^xc (O) NR ^yr ^z,-OR ^xc (O) OR ^y-,-OC (O) R ^x,-OC (O) NR ^xr ^y,-R ^xnR ^yc (O) R ^z,-R ^xoR ^y,-R ^xc (O) OR ^y,-R ^xc (O) NR ^yr ^z,-R ^xc (O) R ^x,-R ^xoC (O) R ^y,-SR ^x,-SOR ^x,-SO ₂r ^xand-ONO ₂, the R wherein in above each group ^x, R ^yand R ^zcan be hydrogen, replace or unsubstituted alkyl, replace or unsubstituted alkoxyl, replace or unsubstituted thiazolinyl, replace or unsubstituted alkynyl, replace or unsubstituted aryl, replace or unsubstituted aryl alkyl, replace or unsubstituted cycloalkyl, replace or unsubstituted cycloalkyl-alkyl, replace or unsubstituted cycloalkenyl group, replace or unsubstituted heteroaryl, replace or unsubstituted heteroaryl alkyl, replace or unsubstituted heterocycle, that replace or unsubstituted cycloheteroalkylalkyl ring or replacement or unsubstituted amino, or Rx, in Ry and Rz arbitrarily both can engage to form replacement or unsubstituted saturated or undersaturated 3-10 person's ring, described ring optionally comprises and may be the same or different and be selected from O, NR ^f(wherein R ^ffor hydrogen or replacement or unsubstituted alkyl) or the hetero atom of S.

23. pharmaceutical compositions according to claim 19, wherein said PI3K δ inhibitor is selected from the group of the following composition:

And its pharmaceutically acceptable salt.

24. pharmaceutical compositions according to claim 19, wherein said dual PI3K δ and gamma inhibitors are selected from the group of the following composition:

And its pharmaceutically acceptable salt.

25. according to claim 19 to the pharmaceutical composition according to any one of 24, and wherein said PDE-4 inhibitor is selected from the group of the following composition: enprofylline, theophylline, aminophylline, Oxtriphylline, A Pusite, roflumilast, cilomilast, appropriate Fei Site, pumafentrine, Li meter Si Te, arofylline, Ah 's azoles is blue, Ao Ge meter Tan, D-4418, Bay-198004, BY343, CP-325,366, D-4396 (Sch-351591), AWD-12-281 (GW-842470), NCS-613, CDP-840, D-4418, PD-168787, T-440, T-2585, V 11294A, CI-1018, CDC-801, CDC-3052, D-22888, YM-58997, Z-15370, N-(the chloro-1-oxo-pyridin of 3,5-bis--4-base)-4-difluoro-methoxy-3-cyclopropyl-methoxy yl-benzamide, (-)-p-[(4aR*, 10bS*)-9-ethyoxyl-1,2,3,4,4a, 10b-six hydrogen-8-methoxyl group-2-methyl benzo [s] [1,6]-benzodiazine-6-base]-N, N-diisopropylbenzamide, (R)-(+)-1-(4-bromobenzyl)-4-[(3-cyclopentyloxy)-4-methoxyphenyl]-2-Pyrrolidone, 3-(cyclopentyloxy-4-methoxyphenyl)-1-(4-N '-[N-2-cyano-S-methyl-different sulfo-urea groups] benzyl)-2-Pyrrolidone, along [4-cyano group-4-(3-cyclopentyloxy-4-methoxyphenyl) cyclohexane extraction-1-carboxylic acid], 2-carbomethoxy-4-cyano group-4-(3-cyclo propyl methoxy-4-difluoro-methoxy phenyl) cyclohexane extraction-1-ketone, along [4-cyano group-4-(3-cyclo propyl methoxy-4-difluoro-methoxy phenyl) cyclohexane extraction-1-alcohol], acetic acid (R)-(+)-ethyl [4-(3-cyclopentyloxy-4-methoxyphenyl) pyrrolidine-2-subunit] ester, acetic acid (S)-(-)-ethyl [4-(3-cyclopentyloxy-4-methoxyphenyl) pyrrolidine-2-subunit] ester, 9 cyclopenta-5,6-dihydro-7-ethyl-3-(2-thienyl)-9H-pyrazolo [3,4c]-1,2,4-triazol [4,3a] pyridine and 9-cyclopenta-5,6-dihydro-7-ethyl-3-(tert-butyl group)-9H-pyrazolo [3,4c]-1,2,4-triazol [4,3a] pyridine, and its pharmaceutically acceptable salt.

26. according to claim 19 to the method according to any one of 25, and wherein said PDE-4 inhibitor is selected from the group of the following composition: theophylline, aminophylline, Oxtriphylline, roflumilast and A Pusite and pharmaceutically acceptable salt.

27. according to claim 19 to the pharmaceutical composition according to any one of 26, and wherein said compositions comprises the described PI3K δ of 0.01mg to about 1000mg or the described PDE4 inhibitor of dual PI3K δ and gamma inhibitors and 0.01mg to about 1000mg.

28. according to claim 19 to the pharmaceutical composition according to any one of 27, and it is used for the treatment of in the method for following disease: autoimmune, respiratory and/or inflammatory diseases or condition of illness, described disease or condition of illness are selected from the group of the following composition: asthma, chronic obstructive pulmonary disease, rheumatoid arthritis, inflammatory bowel, glomerulonephritis, neuroinflammatory disorder, multiple sclerosis, uveitis, psoriasis, arthritis, vasculitis, dermatitis, osteoarthritis, inflammatory myopathy, allergic rhinitis, vaginitis, interstitial cystitis, scleroderma, osteoporosis, eczema, allograft or xenotransplantation (organ, bone marrow, stem cell and other biological cells and tissues) transplant rejection, graft is to versus-host disease, lupus erythematosus, inflammatory diseases, type i diabetes, pulmonary fibrosis, dermatomyositis, house Glenn syndrome, thyroiditis (struma lymphomatosa and autoimmune thyroiditis), myasthenia gravis, autoimmune hemolytic anemia, cystic fibrosis, chronic recurrent hepatitis, primary biliary cirrhosis, anaphylaxis conjunctivitis and atopic dermatitis and its combination.

29. according to claim 19 to the purposes of the pharmaceutical composition according to any one of 28, and it is for the manufacture of the medicament for the following disease for the treatment of: autoimmune, respiratory and inflammatory diseases and condition of illness, described disease and condition of illness are selected from asthma, chronic obstructive pulmonary disease, rheumatoid arthritis, inflammatory bowel, glomerulonephritis, neuroinflammatory disorder, multiple sclerosis, uveitis, psoriasis, arthritis, vasculitis, dermatitis, osteoarthritis, inflammatory myopathy, allergic rhinitis, vaginitis, interstitial cystitis, scleroderma, osteoporosis, eczema, allograft or xenotransplantation (organ, bone marrow, stem cell and other biological cells and tissues) transplant rejection, graft is to versus-host disease, lupus erythematosus, inflammatory diseases, type i diabetes, pulmonary fibrosis, dermatomyositis, house Glenn syndrome, thyroiditis (struma lymphomatosa and autoimmune thyroiditis), myasthenia gravis, autoimmune hemolytic anemia, cystic fibrosis, chronic recurrent hepatitis, primary biliary cirrhosis, anaphylaxis conjunctivitis and atopic dermatitis and its combination.

30. 1 kinds of test kits being used for the treatment of autoimmune, respiratory or inflammatory diseases or condition of illness, described test kit comprises:

(ii) optionally, for using the instructions of described PI3K δ or PI3K δ and gamma inhibitors and PDE4 inhibitor for treating autoimmune, respiratory or inflammatory diseases or condition of illness; And

31. test kits according to claim 30, wherein said PI3K δ or dual PI3K δ and gamma inhibitors and PDE4 inhibitor are used for the treatment of autoimmune, respiratory or inflammatory diseases or condition of illness, described disease or condition of illness are selected from asthma, chronic obstructive pulmonary disease, rheumatoid arthritis, inflammatory bowel, glomerulonephritis, neuroinflammatory disorder, multiple sclerosis, uveitis, psoriasis, arthritis, vasculitis, dermatitis, osteoarthritis, inflammatory myopathy, allergic rhinitis, vaginitis, interstitial cystitis, scleroderma, osteoporosis, eczema, allograft or xenotransplantation (organ, bone marrow, stem cell and other biological cells and tissues) transplant rejection, graft is to versus-host disease, lupus erythematosus, inflammatory diseases, type i diabetes, pulmonary fibrosis, dermatomyositis, house Glenn syndrome, thyroiditis (struma lymphomatosa and autoimmune thyroiditis), myasthenia gravis, autoimmune hemolytic anemia, cystic fibrosis, chronic recurrent hepatitis, primary biliary cirrhosis, anaphylaxis conjunctivitis and atopic dermatitis.

32. test kits according to claim 30 or 31, wherein said PI3K δ or dual PI3K δ and gamma inhibitors are selected from the compound of formula (I), (II), (IA-I), (IA-II), (IA-III) and (IA-IV).

33. test kits according to claim 30 or 31, wherein said PI3K δ inhibitor is selected from:

And its pharmaceutically acceptable salt.

34. test kits according to claim 30 or 31, wherein said dual PI3K δ and gamma inhibitors are selected from:

And its pharmaceutically acceptable salt.